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Am. J. Hum. Genet. 74:1023-1034, 2004

Origin, Diffusion, and Differentiation of Y-Chromosome Haplogroups E

and J: Inferences on the Neolithization of Europe and Later Migratory

Events in the Mediterranean Area

Ornella  Semino,1  Chiara  Magri,1  Giorgia  Benuzzi,1  Alice  A.  Lin,2  Nadia  Al-Zahery,1,4 Vincenza  Battaglia,1  Liliana  Maccioni,5  Costas  Triantaphyllidis,6  Peidong  Shen,7 Peter  J.  Oefner,7  Lev  A.  Zhivotovsky,8  Roy  King,3  Antonio  Torroni,1  L.  Luca  Cavalli-Sforza,2 Peter  A.  Underhill,2  and A.  Silvana  Santachiara-Benerecetti1 1Dipartimento di Genetica e Microbiologia ‘‘A. Buzzati Traverso,” Universita` di Pavia, Pavia, Italy; Departments of 2Genetics and 3Psychiatry and Behavioral Sciences, University of Stanford, Stanford; 4Department of Biotechnology, College of Science, University of Baghdad, Baghdad; 5Istituto di Clinica e Biologia Evolutiva, Universita` di Cagliari, Cagliari, Italy; 6Department of Genetics, Development and Molecular Biology, Aristotle University of Thessaloniki, Thessaloniki; 7Stanford Genome Technology Center, Palo Alto; and 8Vavilov Institute of General Genetics, Russian Academy of Science, Moscow

The phylogeography of Y-chromosome haplogroups E (Hg E) and J (Hg J) was investigated in 12,400 subjects from 29 populations, mainly from Europe and the Mediterranean area but also from Africa and Asia. The observed 501Hg E and 445 Hg J samples were subtyped using 36 binary markers and eight microsatellite loci. Spatial patterns reveal that (1) the two sister clades, J-M267 and J-M172, are distributed differentially within the Near East, North Africa,  and  Europe;  (2)  J-M267  was  spread  by  two  temporally  distinct  migratory  episodes, the most recent one probably associated with the diffusion of Arab people; (3) E-M81 is typical of Berbers, and its presence in Iberia and Sicily is due to recent gene flow from North Africa; (4) J-M172(xM12) distribution is consistent with a Levantine/ Anatolian dispersal route to southeastern Europe and may reflect the spread of Anatolian farmers; and (5) E-M78 (for  which  microsatellite  data  suggest  an  eastern  African  origin)  and,  to  a  lesser  extent,  J-M12(M102)  lineages would trace the subsequent diffusion of people from the southern Balkans to the west. A 7%-22% contribution of Y chromosomes from Greece to southern Italy was estimated by admixture analysis.

It has been proposed that the observed decreasing frequency gradients of Y-chromosome superhaplogroups E (Hg E) (defined by the SRY4064  mutation) and J (Hg J) (characterized  by  the  12f2a-8kb  allele)  (Semino  et  al. 1996; Hammer et al. 1998; Rosser et al. 2000) reached southwestern Europe as a result of demic expansions of Neolithic agriculturalists from the Middle East (Semino et al. 1996; Hammer et al. 1998). The spatial frequency patterns of Hg E and Hg J, at this level of molecular resolution, accommodate both infiltrations of Neolithic agriculturalists into southwestern Europe and cultural adaptations in western and northern Europe by indigenous Mesolithic peoples. This is consistent with the Neolithic migration   hypothesis  (Ammerman  and  Cavalli-Sforza 1984; Cavalli-Sforza 2002). However, this first-order level of molecular resolution does not readily reflect apparent complexities  in  regional  and  local  archaeological  sequences.  The  archaeological  records  suggest  that the large-scale clinal patterns of Hg E and Hg J reflect a mosaic  of  numerous  small-scale, more regional population movements, replacements, and subsequent expansions overlying previous ranges. The recent findings of many biallelic  markers,  which  subdivide  these  two  haplogroups, give us the opportunity to investigate the contribution of different population movements that have spread Hg E and Hg J. Through analysis of the Alu insertion (YAP), the M174 and SRY4064  mutations, and the 12f2a deletion, we  identified  haplogroups  D  (YAP/M174),  E  (YAP/ SRY4064), and J (12f2a) Y chromosomes in 12,400 males from 29 populations, mainly from Europe and the Mediterranean area but also from Africa and Asia. No subject belonged to the recently reported paragroup DE* (Weale at al. 2003), and only 6 belonged to the Asian-specific Hg D, whereas 501 were members of Hg E and 445 of Hg  J.  The  survey  of  36  biallelic  markers  in  the  Hg  E and Hg J Y chromosomes allowed us to define the phylogenetic relationships of their numerous subclades (figs.1 and 2) and to analyze their distributions in the various geographic areas (tables 1 and 2). In addition, the survey of eight microsatellites (figs. 3 and 4) in a subset of these samples  allowed  investigation  of  the  relative  dating  of different subclades.

Received December 17, 2003; accepted for publication February 6, 2004; electronically published April 6, 2004. Address for correspondence and reprints: Dr. Ornella Semino, Dipartimento di Genetica e Microbiologia, Universita`  di Pavia, Via Ferrata, 1, 27100 Pavia, Italy. E-mail: semino@ipvgen.unipv.it2004 by The American Society of Human Genetics. All rights reserved. 0002-9297/2004/7405-0021$15.00

Figure 1     Phylogeny and frequency distributions of Hg E and its main subclades (panels A-G). The numbering of mutations is according to the Y Chromosome Consortium (YCC) (YCC 2002; Jobling and Tyler-Smith 2003). To the left of the phylogeny, the ages (in 1,000 years) of the boxed mutations are reported, with their SEs (Zhivotovsky et al. 2004). Because the procedure used is based on STR data, it actually estimates the ages of STR variation observed within the corresponding haplogroup in the studied populations. With the exception of the value relative to SRY4064   mutation, which as been calculated as TD   (with V0  p 0 ) between the sister clades Hg E-P2 and Hg E-M33, the other values were estimated as the average squared difference (ASD) in the number of repeats between all current chromosomes of a sample and the founder haplotype, which has an expected value mt for single-step mutations (Thomas et al. 1998) and wt for a general mutation scheme, where w is an  average  effective  mutation  rate  at  the  loci,  taken  as  6.9 # 10  4   per  25  years  (Zhivotovsky  et  al.  2004)  (microsatellite  data  available  on request). In some cases, because of small sample sizes or long time passed since the occurrence of the mutation, the founder haplotype could not be reliably estimated as a modal haplotype. Therefore, we constructed it from modal alleles at single loci, although this can underestimate the age if the candidate founder haplotype differs from the real one. To make the computation of the P2 and M35 ages independent from those of their most-represented subclades, the STR variation observed at only the “asterisk” lineages (e.g., E-P2*) has been used. The M35 estimate is in agreement with those of Bosch et al. (2001) and Cruciani et al. (2004 [in this issue]), obtained with different methods. The YAP insertion was studied as an amplified fragment-length polymorphism (Hammer and Horai 1995). The other mutations were investigated in a hierarchical order by use of the denaturing high-performance liquid chromatography (DHPLC) methodology (Underhill et al. 2001). Subhaplogroups observed in this study are illustrated by continuous lines, whereas subhaplogroups discussed elsewhere are indicated by dotted lines. For simplicity, the prefix  “M”  was  omitted  from  the  name  of  the  marker  mutations.  Haplogroup-frequency  surfaces  were  graphically  computer reconstructed following the Kringing procedure (Delfiner 1976) by use of the Surfer System (Golden Software) and the data reported in table 1.

Hg E (fig. 1A) is observed in Africa, Europe, and the Near East and includes the subhaplogroups E-M33, E- M75, and the most widespread subclade, E-P2. The latter includes three clusters, two of which, E-M2 and E- M35, are the most widespread. Haplogroups E-M33 (fig.1B), E-M75 (fig. 1C), and the not-shown E-P2* and E- M2 are virtually absent in European populations and appear to be geographically restricted to sub-Saharan Africa. The E-P2* lineages were observed mainly in Ethiopians, whereas E-M2, which is considered a signature of the Bantu  expansion  (Hammer et al. 1998; Passarino et al.1998; Scozzari et al. 1999), shows its highest frequency(180%) in Senegal and has been sporadically observed in North Africa and Iraq. E-M35 (fig. 1D) has been found in Africa, the Near East, and Europe, where it is believed to have arrived in Neolithic times (Hammer et al. 1998; Semino  et  al.  2000).  In  particular,  from  among  its subgroups,  E-M78  (fig.  1E)  is present in Europe, the Middle  East,  and  North  and  East  Africa.  However, whereas no preferential YCAII microsatellite motif is observed  in  the  Middle  East,  prevalent  associations with YCAIIa21-YCAIIb19 in Europe and YCAIIa22- YCAIIb19 in Africa are found. E-M81 (fig. 1F) is almost absent in Europe (with the exception of Sicily and Iberia) and  the  Middle  East  but  characterizes  the  majority  of the Y chromosomes of populations from northwestern Africa. E-M123 (fig. 1G) is spread in the Near East and is  also  observed  in  North  Africa  and  Europe  but  does not  reach  the  western  European  regions.  E-M281  and E-M329 are geographically restricted, having been seen only in Ethiopians (two subjects each). The remaining 37 E-M35* Y chromosomes were found mainly in Africa, with a high frequency in the Ethiopians and the Khoisan.

Both phylogeography and microsatellite variance suggest that E-P2 and its derivative, E-M35, probably originated in eastern Africa. This inference is further supported by the presence of additional Hg E lineal diversification and by the highest frequency of E-P2* and E-M35* in the same region. The distribution of E-P2* appears limited  to  eastern  African  peoples.  The  E-M35*  lineage shows its highest frequency (19.2%) in the Ethiopian Oromo but with a wider distribution range than E-P2*. Indeed, it is also found at high frequency (16.7%) in the Khoisan of South Africa (Underhill et al. 2000; Cruciani et al. 2002) (suggesting, once again, their ancient relationship with Ethiopians) and observed in southern Europe  (present  study).  It  is  interesting  that  both  E-P2* and E-M35* and their derivatives, E-M78 and E-M123, exhibit in Ethiopians the 12-repeat allele at the DYS392 microsatellite locus, an allele scarcely seen (Y-Chromo- some  STR  Database),  especially  in  other  haplogroups and other populations (A.S.S.-B., unpublished data). In addition, the Ethiopian DYS392-12 allele is usually associated with the unusually short DYS19-11 allele, which is typical of this area. These findings are not easily explained. One possible scenario is that an ancient differentiation of the E-P2 haplogroup occurred in loco (East Africa). However, this also implies a low mutability of the associated microsatellite motif (DYS392-12/DYS19-11).  Alternatively,  the  microsatellite  motif  may  be  due to homoplasy.

The first scenario is more likely, since this unique microsatellite haplotype occurs in E-P2*, E-M35*, and EM78 but is almost absent in all other haplogroups and populations. In addition, the high stability of the DYS392 locus (Brinkmann et al. 1998; Nebel et al. 2001) and of the shorter alleles of DYS19 (Carvalho-Silva et al. 1999) has been reported elsewhere. Moreover, the observation that  the  derivative  E-M78  displays  the  DYS392-12/ DYS19-11  haplotype  suggests  that  it  also  arose in  East Africa. This is illustrated by the microsatellite network (fig. 3, shaded area), which reveals that the Ethiopian branch  harboring  DYS392-12  is  not  shared with either Near Eastern or European populations. The very low frequency of E-M123 in Ethiopia does not allow any inferences about the origin of this clade. The network of E- M78  and  that  of  E-M123  are  in  agreement  with  the hypothesis of their ancient presence in the Near East and their subsequent expansion into the southern Balkans. The divergence time (TD) (Zhivotovsky 2001) between the Near East and European lineages has been estimated to a range of 7-14 thousand years (ky) ago. Cinniog˘ lu et al. (2004) found a high degree of variance of E-M123 in Turkey, which has been interpreted as being due to multiple  founders  rather  than  a  single  early  dispersal event that has remained geographically circumscribed. E-M81 has the lowest variance and a compact network (fig. 3), indicating  either  its  relatively  recent  origin  followed by expansion or its recent expansion after a bottleneck. In Europe,  this  clade  is  restricted  to  the  southernmost regions, such as Iberia and Sicily, and the absence of microsatellite variation suggests a very recent arrival from North  Africa,  consistent  with  previous  observations (Bosch et al. 2001). The frequency pattern and the microsatellite network of E-M2(xM191) (fig. 3) indicate a West African origin followed by expansion, a result that is in agreement with the findings of Cruciani et al. (2002).

Figure 2     Phylogeny and frequency distributions of Hg J and its main subclades (panels A-F). The numbering of mutations is according to  the  YCC  (YCC  2002;  Jobling  and  Tyler-Smith  2003).  To  the  left  of  the  phylogeny,  the  ages  (in  1,000  years)  of  the  boxed mutations are reported, with their SEs (Zhivotovsky et al. 2004). With the exception of the age relative to the 12f2 mutation, which has been estimated as TD   (with V0  p 0) between the combined data of the two sister clades Hg J-M267 and Hg J-M172, the other values have been determined as ASD, as described in figure 1. The 12f2a marker was examined as an RFLP by Southern blotting (Passarino et al. 1998); the other mutations were investigated in hierarchical order by use of DHPLC methodology (Underhill et al. 2001). Three new mutations, M327, M280, and M390, were found in this study. M327 is a TrC transition at np 404 within the STS containing mutation M92, M280 is a GrA transition at np 330 within the STS containing the mutation M67, and M390 is an A insertion after nt 175 in the STS containing the M365 mutation. Conventions used are the same as for figure 1. The frequency surfaces were drawn using the data reported in table 2 and, for Hg J (panel A), also the data from Rosser et al. (2000), Quintana-Murci et al. (2001), and Scozzari et al. (2001).

The 12f2a mutation, which characterizes haplogroup J, was observed in 445 subjects. Hg J harbors two main clades  (see  phylogeny  in  fig.  2),  J-M267  (Cinniog˘ lu  et al. 2004) and J-M172. J-M172 is the more frequent and currently differentiates into eight subhaplogroups defined by mutations M12/M102, M47, M67/M92, M68, M137, M158, M339, and M340, four of which occur at informative frequencies. The less-heterogeneous clade J-M267 includes all of the other 12f2a Y chromosomes that were reported  elsewhere  as  Eu10  (Semino  et  al.  2000).  Its current  level  of  subdivision  includes  five  scarcely  rep- resented subclusters defined by mutations M62, M365, M367/M368, and M369 (Cinniog˘ lu et al. 2004) and by the new mutation M390. Similar to Hg E, different geo- graphic distributions are displayed by the various sub- haplogroups of J (fig. 2). J-M172 (fig. 2C), which occurs as frequently as J-M267 (fig. 2B) in some Middle Eastern populations, is the more prevalent in Europe. Among its subclades, J-M137, J-M158, J-M339, and J-M340 were reported  elsewhere as  single observations (Underhill et al. 2000; Cinniog˘ lu et al. 2004) and have not been ob- served in this study. Likewise, J-M47 and J-M68 characterize very few Near Eastern and Asian samples. How- ever,  J-M12  and  J-M67  and  their  derivatives  are  in- formative,  being  diffused  in  Europe and  observed also in Asia. J-M12 is almost totally represented by its sub- lineage J-M102, which shows frequency peaks in both the  southern  Balkans and  north-central Italy (fig. 2D). The  history  underlying  this  apparent  affinity  remains uncertain. J-M67 (fig. 2E) includes J-M67* lineages (not shown), which are most frequent in the Caucasus, and J-M92,  which  indicates  affinity  between  Anatolia  and southern  Italy  (fig.  2F).  Finally,  the  J-M172*  lineages display a decreasing frequency gradient from the Near East toward western Europe and strongly contribute to the  overall  gradient  of  Hg  J.  J-M267  is  notable,  since this  haplogroup  shows  its  highest  frequencies  in  the Middle  East,  North  Africa,  and  Ethiopia  (fig. 2B)  and its lowest in Europe, having been observed only in the Mediterranean  area.  Of  its  five  subhaplogroups,  only two have been observed: the J-M365 (in two Turks and one  Georgian)  and  the  new  subclade  J-M390  (in  one Lebanese).

a    Numbers in parentheses indicate the number of Y chromosomes analyzed. The population samples include those reported by Semino et al. (2000, 2002), Passarino et al. (1998), and Al-Zahery et al. (2003).

b    An asterisk (*) indicates chromosomes that belong to a clade but not its subclades.

c    The clade 2* also includes the subhaplogroups classified elsewhere as M116.1, M155 (Underhill et al. 2000), M10, and M149 (Cruciani et al. 2002).

d    Data from Cruciani et al. (2002) and F. Cruciani, personal communication.

e    Data from Bosch et al. (2001).

f    Data from Underhill et al. (2000).

g    Data from Semino et al. (2002).

h    The sample “Calabria 2” refers to the Albanian community of the Cosenza province (Torroni et al. 1990).

a    Numbers in parentheses indicate the number of Y chromosomes analyzed. The population samples include those reported by Santachiara-Benerecetti et al. (1993), Semino et al. (1996, 2000, 2002), Passarino et al. (1998), and Al-Zahery et al. (2003).

b    An asterisk (*) indicates chromosomes that belong to a clade but not its subclades.

c    All  chromosomes  classified  as  J*  (because  of  not  belonging  to  J-M172)  by  Cruciani  et  al.  (2002),  Nebel  et  al.  (2001),  and  Bosch  et  al.

(2001) were considered members of J-M267*.

d    Data from Cruciani et al. (2002).

e    Data from Bosch et al. (2001). These samples were not subclassified and are reported only in the “M172 Total” column.

f    Data from Underhill et al. (2000).

g    Data from Nebel et al. (2001).

h    The sample “Calabria 2” refers to the Albanian community of the Cosenza province (Torroni et al. 1990).

Figure 3     Networks of the STR haplotypes of the main subhaplogroups of Hg E. These networks were obtained by the analysis of a subsetof  the  samples  for  the  following  microsatellites:  YCAIIa,  YCAIIb  (Mathias  et  al.  1994),  DYS19,  DYS389,  DYS390, DYS391, and DYS392 (Roewer  et  al.  1996).  The  phylogenetic  relationships  between the  microsatellite haplotypes were determined using the program NETWORK 2.0b  (Fluxus  Engineering).  Networks  were  calculated  by  the  median-joining  method  (p 0 )  (Bandelt  et  al.  1995),  weighting  the  STR  loci according  to  their  relative  variability  in  Hg  E  and,  with  the  exception  of  E-M81,  after  having  processed  the  data  with  the  reduced-median method. Circles represent the microsatellite haplotypes. Unless otherwise indicated by a number on the pie chart, the area of the circles and the area of the sectors are proportional to the haplotype frequency in the haplogroup and in the geographic area indicated by the color. The smallest circle of each network corresponds to one Y chromosome. The shaded area in E-M78 indicates the branch characterized by the DYS392-12 allele.

The extent of differentiation of Hg J, observed both with  the  biallelic  and  microsatellite markers, points to the Middle East as its likely homeland. In this area, J- M172 and J-M267 are equally represented and show the highest degree of internal variation, indicating that it is most likely that these subclades also arose in the Middle East.  However,  their  different  frequencies  in  different Middle Eastern countries and in Europe suggest distinct demography processes, possibly in population groups that underwent different temporal expansions. This is especially true for J-M172. The majority of its lineages are undifferentiated  and  thus  potentially  paraphyletic  (fig.4). Although J-M172* encompasses most of the M172 Y chromosomes in continental Europe and India (Kivi- sild  et  al.  2003;  present study), their degree of affinity and  shared  history  remain  uncertain.  The  J-M67*,  J- M92,  and  J-M102  representatives reflect more distinctive origins and dispersal patterns. Whereas J-M67* and J-M92 show higher frequencies and variances in Europe (0.40  and  0.32,  respectively)  and  in  Turkey  (0.32  and 0.30,  respectively  [Cinniog˘ lu  et  al.  2004])  than  in  the Middle East (0.17 and 0.09, respectively), J-M12(M102) shows  its  maximum  frequency  in the Balkans. In spite of the relative high value of variance of this haplogroup in Turkey (Cinniog˘ lu et al. 2004)-which, however, could be due to multiple arrivals-the pattern of distribution and  the  network  of  J-M12(M102)  (figs.  2  and  4)  are consistent with its diffusion in Europe from the southern Balkans. On the contrary, J-M67* and J-M92 could have arrived in Europe from Anatolia via the Bosphorus isthmus, as well as by seafaring Neolithic populations who reached  southern  Italy.  J-M67*  and  J-M92  could  rep- resent,  at  least  in  part,  the  Y-chromosome  component that King and Underhill (2002) found to correlate with the  distribution,  from  Anatolia  toward  Europe,  of  archaeological painted pottery and anthropomorphic figurines.  On  the  other  hand,  J-M67-  and  J-M12-related lineages have been observed in Pakistan and India; thus, they probably have marked other migratory events, but the  small  number  of  J  subclades  in  these  regions  (Underhill  et  al.  2000;  Kivisild  et  al.  2003;  present study) does not allow an evaluation of the mode and time of their arrival.Southern Italy (Apulia and Calabria) contains sites of the early Neolithic period (Whitehouse 1968), but we know from history that these regions were subsequently colonized by the Greeks (Peloponnesians). To test the relative contribution of Greek colonists versus putative earlier Neolithic settlers, an admixture analysis (Bertorelle and Excoffier 1998) was performed, using E-M78 and J-M172(xM12)  as  signatures  of  Greek  and  Anatolian lineages, respectively. The Anatolian source population was  based  on  523  Turks,  of  whom  118  were  J- M172(xM12)  and  25  were  E-M78  (Cinniog˘ lu  et  al. 2004).  The  Greek  population  comprised  36  Peloponnesian samples, 5 of which were J-M172(xM12) and 17 of which were E-M78 (R.K., unpublished data). In spite of the small Peloponnesian sample size, the high E-M78 frequency  (47%)  observed  here  is  consistent  with that (44%) independently found in the same region (Di Gia- como et al. 2003) for the YAP chromosomes harboring microsatellite haplotypes (A. Novelletto, personal communication) typical of Hg E-M78 (Cruciani et al. 2004 [in this issue]; present study). The admixture analysis yielded   an   admixture   proportion   from  Greece  of 0.07    0.15 for the Calabrian samples and of 0.22    0.15 for  the  Apulian  samples.  SD  was  determined  by  boot- strapping 1,000 replicates.

Figure 4     Network of the STR haplotypes of the main subhaplogroups of Hg J. These networks were obtained by the analysis of a subset of the samples for the following microsatellites: YCAIIa, YCAIIb (Mathias et al. 1994), DYS388 (Thomas et al. 1999), DYS19, DYS389, DYS390, DYS391, and DYS392 (Roewer et al. 1996), by the same procedures used for Hg E (fig. 3). Apart from the YCAII system in Hg J- M267, which was considered as a stable marker in this haplogroup (see text), the STR loci were weighted according to their relative variabilityin Hg J. The most complex networks, J-M267* and J-M172*, were calculated by the median-joining method (p 0 ) on the preprocessed data with the reduced-median method; the other networks were calculated by using only the reduced-median algorithm. The shaded area in J-M267* indicates the branch characterized by the YCAIIa-22/YCAIIb-22 motif. For the areas of the circles and the sectors, see figure 3. The expansion time of this branch was calculated using TD  (Zhivotovsky 2001), which gives 8.7 and 4.3 ky, respectively, for the earliest and the latest bounds of the expansion time. The former estimate was calculated by using the variance in the number of repeats of the remaining six loci, assuming a variance at the beginning of population separation (V0) equal to zero, and thus gives an upper bound for the TD   (Zhivotovsky 2001). The latter assumes a linear approximation of the within-population variance in repeat scores as a function of time and takes a predicted value of V0  prior to population split; because the linearity can be achieved in a case of infinite population size only and because each survived haplogroup started from one individual and could maintain small size for a long time, the linear approximation overestimates V0  and thus might be considered as a lower bound for divergence times (L.A.Z., unpublished method).

The  TD   of  the  two  sister clades J-M267 and J-M172 was estimated, with V0  p 0, and turned out to be 31.7 ky (see phylogeny in fig. 2). This estimate, however, is not easily interpretable, because such old haplogroups are differently represented in different regions where they probably underwent multiple bottlenecks. The lower internal variance of J-M267 in the Middle East and North Africa, relative to Europe and Ethiopia, is suggestive of two different migrations. In the absence of additional binary polymorphisms allowing further informative subdivision of J-M267, the YCAII microsatellite system provides important insights. The majority of J-M267 Y chromosomes harbor  the  single-banded  motif  YCAIIa22-YCAIIb22 in  the  Middle  East  (170%)  and  in  North  Africa (190%), whereas this association is much less frequent in  Ethiopia  and  only  sporadically  found  in  southern Europe. Considering the distribution of this YCAII single-banded pattern-which, besides the usual stepwise mutational  mechanism,  could  be  due  to  a  stable  mutational event (one locus deletion or a single-nucleotide mutation in the primer sequence)-we suggest that the motif  YCAIIa22-YCAIIb22  potentially  characterizes  a monophyletic clade of J-M267. A comparable situation is observed within Hg I-M170, in which the single-banded and a more-recent diffusion (marked by the YCAIIa-22/YCAIIb-22 motif) of Arab people from the southern part of the Middle East toward North Africa.

Acknowledgments

We are grateful to all the donors for providing blood samples and to the people who contributed to their collection. In particular, we thank Ahmet Arslan, Agnese Brega, and B. Kindar (for samples from Turks); Jaume Bertranpetit and Anne Cam- bon-Thomsen (for samples from Catalans, Basques, and Bearnais); Aiping Liu (for samples from Chinese); J. Garcia-Puche (for  samples  from  Andalusians);  and  Adriana  Grasso  and  F. Pignatelli (for samples from Apulians). We warmly acknowl- edge two anonymous reviewers for their helpful and construc- tive  criticism.  This  research  was  supported  by  Progetto  Fin- alizzato CNR “Beni Culturali” (A.S.S.-B.), National Institutes of  Health  grants  GM28428  and  GM55273  (L.L.C.-S.),  Progetto  MIUR-CNR  Genomica  Funzionale-Legge  449/97  (A.T. and L.L.C.-S),  Fondo  d’ Ateneo  per la  Ricerca dell’ Universita` di Pavia (A.S.S.-B and A.T.), the Italian Ministry of the University’ s Progetti Ricerca Interesse Nazionale 2002 and 2003 (A.T.), and Fondo Investimenti Ricerca di Base 2001 (A.T.).

Electronic-Database Information

The URLs for data presented herein are as follows:

Fluxus  Engineering,  http://www.fluxus-engineering.com (for NETWORK 2.0b)

Y-Chromosome  STR  Database,  http://www.cstl.nist.gov/biotech/strbase/y_strs.htm

References

Al-Zahery N, Semino O, Benuzzi G, Magri C, Passarino G, Torroni A, Santachiara-Benerecetti AS (2003) Y-chromosome and mtDNA polymorphisms in Iraq, a crossroad of the early human dispersal and of post-Neolithic migrations. Mol Phy- logenet Evol 28:458-472

Ammerman AJ, Cavalli-Sforza LL (1984) Neolithic transition and the genetics of populations in Europe. Princeton Uni- versity Press, Princeton, NJ

Bandelt HJ, Forster P, Sykes BC, Richards MB (1995) Mito- chondrial portraits of human populations using median net- works. Genetics 141:743-753

Bertorelle G,  Excoffier L (1998) Inferring admixture propor- tions from molecular data. Mol Biol Evol 15:1298-1311

Bosch E, Calafell F, Comas D, Oefner PJ, Underhill PA, Ber- tranpetit J (2001) High-resolution analysis of human Y-chro- mosome variation shows a sharp discontinuity and limited gene flow between northwestern Africa and the Iberian Pen- insula. Am J Hum Genet 68:1019-1029

Brinkmann  B,  Klintschar  M,  Neuhuber  F,  Hu¨ hne  J,  Rolf  B (1998) Mutation rate in human microsatellites: influence of the structure and length of the tandem repeat. Am J Hum Genet 62:1408-1415

Carvalho-Silva DR, Santos FR, Hutz MH, Salzano FM, Pena SD (1999) Divergent human Y-chromosome microsatellite evolution rates. J Mol Evol 49:204-214

Cavalli-Sforza LL (2002) Demic diffusion as the basic process of human expansions. In: Bellwood P, Renfrew C (eds) Ex- amining  the  farming/language  dispersal  hypothesis.  Mc- Donald  Institute  for  Archaeological  Research,  Cambridge, United Kingdom, pp 79-88

Cinniog˘ lu C, King R, Kivisild T, Kalfog˘ lu E, Atasoy S, Cavalleri GL, Lillie AS, Roseman CC, Lin AA, Prince K, Oefner PJ, Shen P, Semino O, Cavalli-Sforza LL, Underhill PA (2004) Excavating Y-chromosome haplotype strata in Anatolia. Hum Genet 114:127-148

Cruciani  F,  La  Fratta  R,  Santolamazza P,  Sellitto D, Pascone R, Moral P, Watson E, Guida V, Colomb EB, Zaharova B, Lavinha J, Vona G, Aman R, Calı` F, Akar N, Richards M, Torroni A, Novelletto A, Scozzari R (2004) Phylogeographic analysis of haplogroup E3b (E-M215) Y chromosomes re- veal multiple migratory events within and out of Africa. Am J Hum Genet 74:1014-1022 (in this issue)

Cruciani  F,  Santolamazza  P,  Shen  P,  Macaulay  V,  Moral  P, Olckers A, Modiano D, Holmes S, Destro-Bisol G, Coia V, Wallace DC, Oefner PJ, Torroni A, Cavalli-Sforza LL, Scoz- zari R, Underhill PA (2002) A back migration from Asia to sub-Saharan Africa is supported by high-resolution analysis of human Y-chromosome haplotypes. Am J Hum Genet 70:1197-1214

Delfiner P (1976) Linear estimation of non-stationary spatial phenomena. Guarasio M, David M, Haijbegts C (eds) Ad- vanced geostatistics in the mining industry. Dordrecht, Rei- del, pp 49-68

Di Giacomo F, Luca F, Anagnou N, Ciavarella G, Corbo RM, Cresta  M,  Cucci  F,  Di  Stasi  L,  Agostiano  V,  Giparaki  M, Loutradis  A,  Mammi  C,  Michalodimitrakis  EN,  Papola F, Pedicini G, Plata E, Terrenato L, Tofanelli S, Malaspina P, Novelletto A (2003) Clinal patterns of human Y chromosomal diversity  in  continental  Italy  and  Greece  are  dominated  by drift and founder effects. Mol Phylogenet Evol 28:387-395

Hammer MF, Horai S (1995) Y chromosomal DNA variation and  the peopling  of Japan.  Am J  Hum  Genet 56:951-962 (erratum 56:1512)

Hammer MF, Karafet T, Rasanayagam A, Wood ET, Altheide TK, Jenkins T, Griffiths RC, Templeton AR, Zegura SL (1998) Out of Africa and back again: nested cladistic analysis of human Y chromosome variation. Mol Biol Evol 15:427-441

Jobling MA, Tyler-Smith C (2003) The human Y chromosome: an evolutionary marker comes of age. Nat Rev Genet 4:598-612

King R, Underhill PA (2002) Congruent distribution of Neo- lithic painted pottery and ceramic figurines with Y-chromo- some lineages. Antiquity 76:707-714

Kivisild T, Rootsi S, Metspalu M, Mastana S, Kaldma K, Parik J, Metspalu E, Adojaan M, Tolk H-V, Stepanov V, Go¨ lge M, Usanga E, Papiha SS, Cinniog˘ lu C, King R, Cavalli-Sforza LL, Underhill PA, Villems R (2003) The genetic heritage of ear- liest settlers persists in both the Indian tribal and caste populations. Am J Hum Genet 72:313-332

Mathias N, Bayes M, Tyler-Smith C (1994) Highly informative compound haplotypes for the human Y chromosome. Hum Mol Genet 3:115-123

Nebel A, Filon D, Hohoff C, Faerman M, Brinkmann B, Oppenheim  A  (2001) Haplogroup-specific deviation from the stepwise mutation model at the microsatellite loci DYS388 and DYS392. Eur J Hum Genet 9:22-26

Nebel A, Filon D, Weiss DA, Weale M, Faerman M, Oppen- heim  A,  Thomas  MG  (2000)  High-resolution  Y  chromo- some haplotypes of Israeli and Palestinian Arabs reveal geo- graphic substructure and substantial overlap with haplotypes of Jews. Hum Genet 107:630-641

Nebel A, Landau-Tasseron E, Filon D, Oppenheim A, Faerman M  (2002)  Genetic  evidence  for  the  expansion  of  Arabian tribes into the southern Levant and North Africa. Am J Hum Genet 70:1594-1596

Passarino G, Semino O, Quintana-Murci L, Excoffier L, Ham- mer M, Santachiara-Benerecetti AS (1998) Different genetic components  in  the  Ethiopian  population,  identified  by mtDNA and Y-chromosome polymorphisms. Am J Hum Ge- net 62:420-434

Quintana-Murci  L,  Krausz  C,  Zerjal  T,  Sayar  SH,  Hammer MF, Mehdi SQ, Ayub Q, Qamar R, Mohyuddin A, Radha- krishna U, Jobling MA, Tyler-Smith C, McElreavey K (2001) Y-chromosome  lineages  trace  diffusion  of  people  and  lan- guages in southwestern Asia. Am J Hum Genet 68:537-542

Roewer L, Kayser M, Dieltjes P, Nagy M, Bakker E, Krawczak M,  de  Knijff  P  (1996)  Analysis  of  molecular  variance(AMOVA) of Y-chromosome-specific microsatellites in two closely related human populations. Hum Mol Genet 5:1029-1033

Rosser  ZH,  Zerjal  T,  Hurles  ME,  Adojaan  M,  Alavantic  D, Amorim A, Amos W, et al (2000) Y-chromosomal diversity in Europe is clinal and influenced primarily by geography, rather than by language. Am J Hum Genet 67:1526-1543

Santachiara-Benerecetti AS, Semino O, Passarino G, Torroni A, Brdicka R, Fellous M, Modiano G (1993) The common Near- Eastern origin of Ashkenazi and Sephardi Jews supported by Y-chromosome similarity. Ann Hum Genet 57:55-64

Scozzari R, Cruciani F, Pangrazio A, Santolamazza P, Vona G, Moral  P,  Latini  V,  Varesi  L,  Memmi  MM,  Romano V,  De Leo G, Gennarelli M, Jaruzelska J, Villems R, Parik J, Ma- caulay V, Torroni A (2001) Human Y-chromosome variation in the western Mediterranean area: implications for the peo- pling of the region. Hum Immunol 62:871-884

Scozzari R, Cruciani F, Santolamazza P, Malaspina P, Torroni A, Sellitto D, Arredi B, Destro-Bisol G, De Stefano G, Rick- ards O, Martinez-Labarga C, Modiano D, Biondi G, Moral P, Olckers A, Wallace DC, Novelletto A (1999) Combined use  of  biallelic  and  microsatellite Y-chromosome polymor- phisms to infer affinities among African populations. Am J Hum Genet 65:829-846

Semino O, Passarino G, Brega A, Fellous M, Santachiara-Be- nerecetti AS (1996) A view of the Neolithic demic diffusion in Europe through two Y chromosome-specific markers. Am J Hum Genet 59:964-968

Semino O, Passarino G, Oefner PJ, Lin AA, Arbuzova S, Beck- man LE, De Benedictis G, Francalacci P, Kouvatsi A, Lim- borska S, Marcikiae M, Mika A, Mika B, Primorac D, Santa- chiara-Benerecetti AS, Cavalli-Sforza LL, Underhill PA (2000) The  genetic  legacy  of  Paleolithic  Homo  sapiens  sapiens  in extant Europeans: a Y chromosome perspective. Science 290:1155-1159

Semino O, Santachiara-Benerecetti AS, Falaschi F, Cavalli-Sforza LL, Underhill PA (2002) Ethiopians and Khoisan share the deepest clades of the human Y-chromosome phylogeny. Am J Hum Genet 70:265-268

Thomas MG, Bradman N, Flin HM (1999) High throughput analysis of 10 microsatellite and 11 diallelic polymorphisms on the human Y-chromosome. Hum Genet 105:577-581

Thomas MG, Skorecki K, Ben-Ami H, Parfitt T, Bradman N, Goldstein DB (1998) Origins of Old Testament priests. Na- ture 394:138-140

Torroni  A, Semino  O,  Rose G,  De Benedictis G, Brancati C, Santachiara Benerecetti AS (1990) Mitochondrial DNA poly- morphisms in the Albanian population of Calabria (southern Italy). Int J Anthropol 5:97-104

Underhill PA, Passarino G, Lin AA, Shen P, Mirazon Lahr M, Foley RA, Oefner PJ, Cavalli-Sforza LL (2001) The phylo- geography of Y chromosome binary haplotypes and the ori- gins of modern human populations. Ann Hum Genet 65:43-62

Underhill PA, Shen P, Lin AA, Jin L, Passarino G, Yang WH, Kauffman E, Bonne-Tamir B, Bertranpetit J, Francalacci P, Ibrahim  M,  Jenkins  T,  Kidd  JR,  Mehdi  SQ,  Seielstad MT, Wells RS, Piazza A, Davis RW, Feldman MW, Cavalli-Sforza LL, Oefner PJ (2000) Y chromosome sequence variation and the history of human populations. Nat Genet 26:358-361

Weale ME, Shah T, Jones AL, Greenhalgh J, Wilson JF, Nyma- dawa  P,  Zeitlin  D,  Connell  BA,  Bradman  N, Thomas M (2003) Rare deep-rooting Y chromosome lineages in humans: lessons for phylogeography. Genetics 165:229-234

Whitehouse  R  (1968)  The  early  Neolithic  of  southern  Italy. Antiquity 42:188-193

Y Chromosome Consortium (YCC) (2002) A nomenclature sys- tem for the tree of human Y-chromosomal binary haplo- groups. Genome Res 12:339-348

Zhivotovsky LA (2001) Estimating divergence time with use of microsatellite genetic distances: impacts of population growth and gene flow. Mol Biol Evol 18:700-709

Zhivotovsky LA, Underhill PA, Cinniog˘ lu C, Kayser M, Morar B, Kivisild T, Scozzari R, Cruciani F, Destro-Bisol G, Spedini G, Chambers GK, Herrera RJ, Yong KK, Gresham D, Tour- nev I, Feldman MW, Kalaydjieva L (2004) The effective mu- tation rate at Y chromosome short tandem repeats, with ap- plication to human population-divergence time. Am J Hum Genet 74: 50-61 Am. J. Hum. Genet. 74:1023-1034, 2004