Polymorphic Alu Insertions and the Asian Origin of Native American Populations

Author(s): GABRIEL E. NOVICK, CORINA C. NOVICK, JUAN YUNIS, EMILIO YUNIS,

PAMELA ANTUNEZ DE MAYOLO, W. DOUGLAS SCHEER, PRESCOTT L. DEININGER, MARK
STONEKING, DANIEL S. YORK, MARK A. BATZER and RENE J. HERRERA
Source: Human Biology, Vol. 70, No. 1 (February 1998), pp. 23-39
Published by: Wayne State University Press
Stable URL: http://www.jstor.org/stable/41465617
Accessed: 15-11-2017 02:10 UTC

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Polymorphie Alu Insertions and the Asian Origin of Native
American Populations

GABRIEL E. NOVICK,1 CORINA . NOVICK,1 JUAN YUNIS,2 EMILIO YUNIS,2

PAMELA ANTUNEZ DE MAYOLO,' W. DOUGLAS SCHEER,3
PRESCOTT L. DEININGER,4 MARK STONEKING,5 DANIEL S. YORK,6
MARK A. BATZER,3 AND RENE J. HERRERA1

Abstract A rapid PCR-based assay was used to study the distribution

of 5 polymorphic Alu insertions in 895 unrelated individuals from 30
populations, 24 from North, Central, and South America. Although a
significant level of interpopulation variability was detected, the variabil-
ity was less than that observed in a worldwide population survey. This
is consistent with the bottleneck effect and genetic drift forces that may
have acted on the migrating founder groups. The results corroborate the
Asian origin of native American populations but do not support the
multiple-wave migration hypothesis supposedly responsible for the tri-
partite Eskaleut, Nadene, and Amerind linguistic groups. Instead, these
populations exhibit three major identifiable clusters reflecting geographic
distribution. Close similarity between the Chinese and native Americans
suggests recent gene flow from Asia.

A wealth of anthropological, dental, linguistic, and genetic data have been

accumulated in the area of human evolution in the Americas. However, the
issues of origin, point of entry, number of migrations, timing, routes of ex-
pansion, and survivorship of native Americans remain controversial. Tradi-
tional anthropological analyses support the hypothesis that native Americans
are derived from northern Asian ancestors who reached the New World by

1 Department of Biological Sciences, Florida International University, University Park Campus, Mi-
ami, FL 33199.
2 Instituto de Gentica, Universidad Nacional de Colombia, Santa F de Bogot, Colombia.
3 Department of Pathology, Stanley S. Scott Cancer Center, Louisiana State University Medicai
Center, 1901 Perdido Street, New Orleans, LA 70112.
4 Department of Biochemistry and Molecular Biology and Center for Molecular and Human Genetics,
Louisiana State University Medical Center, 1901 Perdido Street, New Orleans, LA 70112; and Laboratory
of Molecular Genetics, Alton Ochsner Medical Foundation, New Orleans, LA 70121.
5 Department of Anthropology, Pennsylvania State University, University Park, PA 16802.
6 Department of Natural and Health Sciences, Barry University, 1 1300 N.E. 2d Avenue, Miami, FL
33161.
Address correspondence to R.J. Herrera.

Human Biology, February 1998, v. 70, no. 1, pp. 23-39.

Copyright 1998 Wayne State University Press, Detroit, Michigan 48201-1309

KEY WORDS: ALU INSERTION, NATIVE AMERICANS, PEOPLING OF THE AMERICAS

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24 / NO VICK ET AL.

walking over the Bering land bridge that was exposed during the last glaci-
ation, some 20,000 years ago (Fladmark 1983).
Several hypotheses have been formulated to explain the origins of hu-
mans in the New World. The tripartite hypothesis proposes three waves of
migration that originated in northern Asia and gave rise to the three main
genetic and linguistic clusters: Eskaleut, Nadene, and Amerind (Greenberg
1987). Under this hypothesis there were three distinct migrations. The first
one, sometime before 15,000 years ago, originated the Amerind cluster, which
spread over most of the New World. The second migration, 15,000-10,000
years ago, gave rise to the Nadene group. The third wave, 10,000 years ago,
founded the Eskaleut cluster. This hypothesis has been corroborated by dental
data (Greenberg et al. 1986) and by nuclear and mitochondrial DNA genetic
data (Wallace and Torroni 1992; Cavalli-Sforza et al. 1994).
Multiple migrations also have been proposed (Cavalli-Sforza et al.
1994), as have single-migration models (Rogers et al. 1991). One possibility,
according to the single-migration model, portrays a mostly ice-covered North
America during the last glacial maximum with some ice areas suitable for
habitation (Rogers et al. 1991). The isolation that may have occurred in this
ice-free refugium may have encouraged differentiation, genetic drift, and in-
dependent differentiation (Rogers et al. 1991). This differentiation may have
yielded the profile of multiple migrations postulated by other researchers.
Several polymorphic genetic systems have been used to study native
American phylogeny. These include mitochondrial DNA, variable number of
tandem repeats (VNTRs), restriction fragment length polymorphisms
(RFLPs), and point mutations in Alu sequences (O'Rourke et al. 1992; Schan-
field 1992; Kidd, Pakstis et al. 1993; Torroni et al. 1994; Kidd and Kidd
1996; Knight et al. 1996). Alu sequences are the largest family of short in-
terspersed repetitive elements (SINEs) in humans, with an excess of 500,000
copies per haploid genome [for reviews see Deininger and Batzer (1993) and
Novick et al. (1996)]. Alu elements are ancestrally derived from the endo-
plasmic reticulum signal recognition particle 7SL RNA gene (Ullu and
Tschudi 1984), with which they share about 90% sequence similarity through-
out most of their sequences (Ullu and Tschudi 1984). Alu elements are distrib-
uted throughout the genomes of primates. Recently, one Alu subfamily was
found to be largely human specific (HS) (Batzer 1990; Batzer and Deininger
1991). Members of this subfamily have been inserted recently into the human
lineage genome, within the last 200,000 to 6 million years (Batzer 1991 ; Batzer
and Deininger 1991). A limited number of Alu elements are transcriptionally
active (Matera et al. 1990) and undergo amplification into other genomic lo-
cations (Wallace et al. 1991) in a process termed retroposition.
In addition to the polymorphic nature of many Alu insertions, several
features make Alu elements exceptional genetic markers: the stability of the
Alu insertion event, the lack of a known mechanism for the precise removal
of Alu elements from their specific chromosomal site of insertion, and the

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 Polymorphie Alu Insertions / 25

low rate of de novo insertions that reach polymorphic levels ( atzer and
Deininger 1991). These characteristics make it highly unlikely that the same
Alu insertion could occur more than once independently at the same locus or
that once inserted, an element could be removed without leaving vestiges of
its existence behind. Furthermore, the ancestral state of an Alu insertion in-
variably is the absence (complete and exact) of the element at a particular
locus and the presence of an insertion at that site, the forward mutational
change. This is an invaluable attribute not present in other polymorphic sys-
tems (e.g., RFLPs) where the ancestral state is ambiguous.
We recently reported the application of a polymerase chain reaction
(PCR) based Alu insertion polymorphism assay to the analysis of population
relationships in a worldwide survey (Batzer et al. 1994; Novick et al. 1995).
In the present study we analyzed a group of 24 native American populations
using 5 polymorphic Alu insertion sites with regard to their genetic structure,
interpopulation affinities, and relationships with respect to 6 non-American
groups. The results presented here corroborate the Asian origin of native
American populations but do not support the multiple-migration hypothesis
for the establishment of the tripartite Eskaleut, Nadene, and Amerind lin-
guistic groups. Instead, the phylogenetic relationships among the native
American groups reflect their geographic distribution. The presented data also
demonstrate close phylogenetic relationships between mainland Chinese and
native Americans, particularly Mayan groups. This may indicate recent gene
flow from Asia to the New World.

Materials and Methods

DNA Samples. Twenty-four native American populations were studied.

Their names and origins are provided in Table 1. Their geographic location
are illustrated in Figure 1. The Campeche Maya, Kartiana, Surui, Moskoke
and Quechua samples were kindly provided by Judy R. Kidd [Kidd, Paksti
et al. (1993) and unpublished data].

PCR Amplification. DNA from peripheral blood mononuclear cells o

cells in culture were isolated according to the method of Ausabel et al. (1987).
Five different pairs of primers, each complementary to a different Alu inser-
tion located on different chromosomes, were used: ACE , TPA25 , PV92 , APO,
and F13B. PCR primer sequences, location, and optimal annealing tempera-
tures have been reported previously (Batzer and Deininger 1991; Tiret et al
1992; Batzer et al. 1994; Kass et al. 1994). PCR amplification was carried
out in a final volume of 100 fi' , as described previously (Batzer et al. 1991)
Ten microliters of the PCR products were electrophoresed in 3% IX TAE
agarose gels with 0.5 jug of Hae III digested 174 DNA as a fragment
length marker. Genotypes were scored after staining with ethidium bromide.

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26 / NOVICK ET AL.

Table 1. Names and Origins of the 24 Studied Native American Populations

Population Localization
1 Alaska native Aleut Islands
2 Greenland native Greenland

3 Cree-Ojibwa Central Canada

4 Navajo Southwestern United States
5 Zuni Southwestern United States
6 Sioux North-central United States
7 Moskoke Southeastern United States

8 Campeche Maya Campeche Province, Mexico

9 Buctzotz Maya Yucatan Peninsula, Mexico
10 Waunana Eastern Panama

1 1 Ngobe Western Panama

12 Paez Cauca, southeastern Colombia
13 Guambiano Cauca, southeastern Colombia
14 Ingano Nario, southeastern Colombia
15 Kogui Northern Colombia
16 Arhuacoa Northern Colombia
17 Chimila Northern Colombia
18 Wayuu Northern Colombia
19 Guayabera East-central Colombia
20 Inca Quito, Ecuador
21 Kantiana Amazonia, Brazil
22 Surui Amazonia, Brazil
23 Toba Northwestern Argentina
24 Quechua3 Northern Bolivia
25 Nigerian3 Nigeria, Africa
26 Pygmy3 Zaire, Africa
27 Chinese3 Mainland China
28 Turkish Cypriot3 Cyprus Island
29 Greek Cypriot3 Cyprus Island
30 European American3 United States, European ancestry
a. Populations previously published for some or all of the polymorphic Alu insertions report
study.

Amplified DNA fragments were visualized using ultraviolet light and a Cy-
bertech CSI Imaging System.

Data Analysis. Allele frequencies, heterozygosity, and standard errors

were calculated for all five loci, and the Hardy-Weinberg expected number
was calculated for each genotype. Chi-square tests for goodness of fit were
performed. Average heterozygosity, the associated standard error, and GST
values were determined according to equations previously reported (Nei
1987). GST values are a measure of the proportion of the total genetic variance
that can be attributed to between-population differences.
Genetic distance analysis was performed using the simdis routine in the
biosys program (D.L. Swofford and R.B. Selander, University of Illinois,

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 Polymorphie Alu Insertions / 27

Figure 1. Geographie distribution of the native American populations examined in this study. See
Table 1 for the names of the numbered populations.

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28 / NO VICK ET AL.

Champaign-Urbana) according to the methods of Reynolds et al. (1983), Nei

(1972), and Cavalli-Sforza and Edwards (1967). Each distance matrix was
used to construct a tree by means of the unweighted pair group method using
arithmetic means (upgma) (data not shown).
Maximum-likelihood analyses were performed using the contml pro-
gram. The relative amount of gene flow in each population was calculated by
plotting the heterozygosity of each population against the distance of the
population from the centroid, as described by Harpending and Ward (1982).
The distance from the centroid r for a population / is
r, = (Pi - Pf/P(l - p ), (1)
where pt and P are the frequencies of the A
the total population, respectively. The th
should be a linear relationship between hete
centroid. Populations that have experienced
above the theoretical prediction, whereas
less gene flow than average fall below the

Results

Genetic Variation within Populations. The distribution of 5 polymorphic

Alu insertions was determined in 895 unrelated individuals from 30 popula-
tions. From that sample 691 individuals belonged to 24 native American
populations and the remaining 204 individuals belonged to the 6 reference
groups (Chinese, N = 31; Nigerians, N = 11; African Pygmies, N = 34;
Turkish Cypriots, N= 33; Greek Cypriots, N = 50; Americans of European
ancestry, N = 45) from geographic locations outside North, Central, or South
America. The allele frequencies and heterozygosity for each locus are re-
ported in Table 2.
All Alu insertions were polymorphic in all native American populations
except two. The PV92 locus was fixed for the presence of the insertion in the
Chimila and the APO insertion allele was fixed in the Chimila, Kantiana,
Sioux, Moskoke, Arhuaco, Kogui, Quechua, Guambiano, Paez, Ingano,
Guayabera, Waunana, Navajo, and Zuni.
One hundred fifty chi-square tests for goodness of fit to Hardy-Weinberg
equilibrium were performed, and 10 tests were significant (Wayuu for TPA25 ,
PV92 , and F13B ; Toba for TPA25 ; Cree-Ojibwa for ACE; Inca for APO and
F13B; native Alaskans from the Aleut Islands for PV92; and native Green-
landers for PV92 and F13B).
The heterozygosity of each population, averaged across the five Alu
insertions, ranged from 0.15 in the Guayabera to 0.45 in the Inca. The het-
erozygosity of each marker, averaged across all the populations, ranged from
0.07 for APO to 0.5 for TPA25. These numbers are quite high, considering
that each of the Alu insertions represents a biallelic polymorphism with a
maximum heterozygosity of 0.5.

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 Polymorphie Alu Insertions / 29

Genetic Differences and Affinity among Populations. Genetic differ-

entiation among populations was estimated by calculating GST values for each
Alu repeat. GST values (Nei 1987) are a measure of the proportion of the total
genetic variance that can be attributed to between-population differences. The
GST estimates ranged from 0.04 for F13B to 0.16 for PV92. All GST values
were significantly different from 0, as determined by chi-square analysis of
the allele frequencies. These results reflect significant differences between the
native American populations with respect to their allele frequencies.
To examine the evolutionary relationships among the populations, we
estimated genetic distances directly from the allele frequencies given in Table
2. The maximum-likelihood is presented in Figure 2. A hypothetical ancestral
population, fixed for the absence of the insertion in all five loci, was placed
in the tree. This analytical strategy was based on previous reports that show
that all five Alu repeats studied here are absent from the genomes of nonhu-
man primates (Batzer and Deininger 1991; Batzer et al. 1994). It is also based
on the facts that the direction of mutation for Alu elements is the insertion
event and that the mechanisms for the entire and exact removal of an element
from its locus are unknown (Sawada and Schmid 1986; Bailey and Shen
1993). The hypothetical ancestral population grouped closer to the African
populations in all phylogenetic trees (e.g., Figure 2). All reference populations
(non- American) grouped outside the native American cluster, except for the
Chinese, which grouped with the native American populations. The Inca are
the native American population that consistently groups away from the native
American group. Coincidentally, the Inca have the highest heterozygosity
(0.45). The native American populations segregated into three groups. The
first, going from top to bottom in Figure 2, predominantly consists of North
American populations [Moskoke, Sioux, Greenland Eskimo, Alaskan natives
(Eskaleut), Buctzotz Maya, Campeche Maya, and Cree-Ojibwa]. The two
Central American populations examined in this study (the Ngobe and the
Waunana) segregated into a paraphyletic group (Figure 2). The third group
consists of populations in a clade made up mostly of South American tribes
(Guambiano, Kogui, Quechua, Ingano, Paez, Guayabera, Chimila, and Kar-
itiana). The Chinese cluster with the North American populations close to the
two Mayan groups.

Gene Flow in Native American Populations. The relative amount of

gene flow for each population was estimated by plotting the heterozygosity
of each group against the genetic distance from the centroid (Harpending and
Ward 1982). This theory assumes a simple linear relationship between the
heterozygosity of a population and the genetic distance of the population from
the centroid (the overall mean allele frequency of the populations). If a popu-
lation is receiving genes from elsewhere at a higher than average rate, then
the heterozygosity will be higher than predicted. A lower than predicted het-
erozygosity implies that the population is more isolated, thereby receiving

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30 / NOVICK ET AL.

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 Polymorphie Alu Insertions /31

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32 / NOVICK ET AL.

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 Polymorphie Alu Insertions / 33

less gene flow than average. Our analysis indicates that the non-American
populations are outliers in terms of having higher than predicted heterozy-
gosity. The exception to this is the Chinese, which group within the native
American populations (Figure 3). Within the native American groups both
the Inca and the Moskoke have heterozygosity values beyond the theoretical
expectation. This finding is consistent with the high average heterozygosity
observed in both groups and the clustering pattern observed for the Inca in
the maximum-likelihood tree.

Discussion

The Alu family of repetitive elements first appeared in primate genomes

about 65 million years ago (Deininger and Daniels 1986). The elements are
still undergoing retroposition by means of an RNA intermediary from Alu
sources or master genes (Deininger and Slagel 1988; Deininger et al. 1992) at
a rate of approximately 100 elements that reach polymorphic levels per mil-
lion years (Batzer et al. 1990; Batzer and Deininger 1991). Alu insertions are
distributed fairly randomly (Daniels and Deininger 1985; Batzer et al. 1991)
and represent stable events that are rarely deleted without leaving behind
traces of their presence (Sawada and Schmid 1986; Bailey and Shen 1993). In
contrast to other multilocus polymorphisms, such as some RFLPs, offspring
inherit from their parents the Alu insertion and lack of insertion alleles as
codominant alleles by simple Mendelian segregation (Novick et al. 1994).
We recently reported the application of Alu insertion polymorphisms to
the study of population phylogenetic relationships in a worldwide analysis of
16 groups (Batzer et al. 1994). In the present study we report the allele dis-
tribution of 5 polymorphic Alu insertions among 691 individuals from 24
native American populations from Alaska to Argentina. Consistent with our
previous report (Batzer et al. 1994), all 5 loci were highly variable with popu-
lation average heterozygosity values of up to 0.45 (Inca) and a maximum
heterozygosity of 0.5 for TPA25.
Ten chi-square tests for goodness of fit to Hardy- Weinberg equilibrium
were significant (Wayuu for TPA25 , PV92 , and F13B' Toba for TPA25' Cree-
Ojibwa for ACE; Inca for APO and F13B; Alaskan natives for PV92 ; and
Greenland Eskimo for PV92 and F13B). This is slightly higher than the 5%
(7.5 significant chi-square tests) expected from chance alone. Furthermore,
all significant values were due to a deficiency of heterozygotes where, if they
were normal statistical fluctuations, a similar number of departures from the
expected number of heterozygotes for excess and deficiency would be ex-
pected. Therefore we conclude that the deficiency in heterozygotes is real and
possibly the reflection of a variable degree of inbreeding within the popula-
tions or a lack of gene flow.
A significant amount of interpopulation variability was observed, with
GST values ranging from 0.04 for F13B to 0.16 for PV92. However, these

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34 / NOVICK ET AL.

Figure 3. Centroid analysis of gene flow. The heterozygosity of each population (_y axis) is plotted
against the distance of the population from the centroid (the overall mean allele fre-
quencies of the populations) (x axis). The upper plot includes all 30 populations, and
the lower plot illustrates the centroid analysis of only the native American populations.
Filled circles represent native American populations; open circles indicate non- American
groups.

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 Polymorphie Alu Insertions / 35

values are smaller than what we observed for a set of worldwide populations
(Batzer et al. 1994), as would be expected from a more closely related set of
populations in the native American group. Aborigines in the New World are
thought to derive from a limited number of individuals. Under these condi-
tions genetic diversity is expected to be reduced because of a bottleneck
effect. In addition, as humans migrated south from Alaska to Tierra del Fuego,
genetic drift would be expected to increasingly limit variability. When the
levels of heterozygosity are examined, we observe that, although the average
heterozygosity among North American groups is 51% higher than the com-
bined Central and South American average heterozygosity, the South Ameri-
can population exhibits an 18% higher average heterozygosity than the Cen-
tral American groups. Although admixture may be responsible for high levels
of heterozygosity in some North and South American populations, the higher
level of heterozygosity in South American groups compared with Central
American populations does not support only north to south migrations of
people from northern Asia to the southernmost regions of South America.
The relationship between populations can be deduced from the topology
of the maximum-likelihood tree (Figure 2). In the tree and consistent with
previous reports (Batzer et al. 1994; Novick et al. 1995), the African groups
are closer to the hypothetical ancestor. This is compatible with a probable
placement of the origin of the polymorphic Alu insertions and of the ancestors
of modern human populations in Africa.
The Inca are found at a distance from the other native American pop-
ulations. In addition, the Inca and Moskoke groups show the highest average
heterozygosity (0.45 and 0.42, respectively) and the highest gene flow (cen-
troid analysis, Figure 3). These results lead us to conclude that these two
populations have experienced a significant degree of admixture with non-
American native groups (i.e., Europeans and/or Africans). It is possible that
at the sites where our samples were collected, these two populations are rela-
tively admixed.
In two cases, the Buctzotz Maya/Campeche Maya and the Alaska/
Greenland natives, closely related populations collected from different
regions cluster together in the maximum-likelihood tree (Figure 2). This also
reflects the specificity and resolving power of polymorphic Alu insertions. It
is significant that the related Alaskan native (Eskaleut) and Greenland Eskimo
populations, located approximately 3000 miles apart, group so closely in the
maximum-likelihood analysis.
Although the three linguistic clusters are not equally represented in our
population set, the maximum-likelihood analysis does not show a distribution
compatible with the Eskaleut (Alaskan and Greenland natives), Nadene (Nav-
ajo), and Amerind (the rest of the native American populations in this study)
linguistic clusters. Rather, the phylogenetic tree exhibits three major identi-
fiable groups reflecting geographic distribution. One group (going from north
to south) includes populations from North America (Sioux, Moskoke, Alas-

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36 / NOVICK ET AL.

kan natives, Greenland natives, Buctzotz Maya, and Campeche Maya). This
cluster also incorporates the Wayuu from Colombia. This tribe has kept a
centuries-old commercial relationship with the populations from the Dutch
Antilles, with which they show a strong degree of admixture (Yunis et al.
1994). This may explain their location in the North American paraphyletic
group, closer to European Americans. The second paraphyletic group includes
populations from Central America (Ngobe and Waunana). The third group
contains most of the populations from South America: Guambiano, Kogui,
Quechua, Ingano, Paez, Guayabero, Chimila, and Kantiana. It is significant
that the Asian (Chinese) population is phylogenetically closer to most of the
North American groups than to the Central and South American groups. Also,
the phylogenetic affinities among native American populations parallel the
north to south geographic distribution of the groups, with most of the South
American populations and North American populations being more related
to the Central American groups than to each other. It is possible that a single
migration to the New World followed by partial isolation and genetic drift
gave rise to this geographic distribution and the observed differences in allele
frequencies between the native American populations.
As previously reported for 42 populations worldwide with 120 allele
frequencies from classical genetic markers (Cavalli-Sforza et al. 1994), Asian
populations, including native American groups, and European populations
cluster on the same branch when analyzed using the 5 polymorphic Alu
insertion loci. Also, as with the classical markers studied by Cavalli-Sforza
et al. (1994), the African populations group by themselves (Figure 2). The
close correspondence between the results from these two types of DNA
marker systems adds support to the significance of /-derived phylogenetic
analysis.
Our analyses also show that the Chinese consistently cluster within the
North American group. This parallels previously published data on mito-
chondrial DNA that establish native American mitochondrial DNA as derived
from a few mitochondrial haplotypes found in Asian populations of south-
eastern China (Shurr et al. 1990; Ballinger et al. 1992; Torroni et al. 1992;
Wallace and Torroni 1992). Yet it contrasts with the position of the Chinese
in phylogenetic trees derived from 120 allele frequencies (Cavalli-Sforza et
al. 1994). Using classical genetic markers, Cavalli-Sforza et al. (1994) found
that the Chinese are located on a different branch from the native Americans,
off the second bifurcation. Eurasian populations, including native Americans,
northeastern Asians, and Caucasoids, group on another branch off the second
bifurcation. Because of their attributes as genetic markers, the polymorphic
Alu insertions may provide a level of phylogenetic sensitivity that allows for
the detection of some evolutionary relationships (Chinese-native American).
Modern humans are assumed to have migrated from Africa to Europe
and Asia and from there to the Americas through the Bering Strait. In this
scenario it should be expected that the Chinese would be located somewhere

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 Polymorphie Alu Insertions / 37

in between the African groups and the native American populations. There-
fore it is interesting that the Chinese group closer to the Amerinds, such as
the Mayan populations, than to Alaskan and Greenland natives, who represent
a more recent and continuing wave of migrants from Asia.
Under conditions where genetic differences are not generated by new
alleles, as in the case of the Alu insertions of this study (all five insertions
took place before humans migrated out of Africa), all genetic differences at
these loci must be the result of genetic drift and possibly selection. Under
these conditions native Americans would be expected to be more different
from each other and from their Asian ancestors. Although genetic drift was
supposedly a prominent phenomenon as humans migrated south in limited
numbers, humans were subjected to ever decreasing diversity, as opposed to
the more open and interactive migrations such as the ones from Africa to Asia
or Europe. In other words, because variations between all human populations,
as far as these five Alu insertions are concerned, are due to bottleneck events
and genetic drift, situations that favor these two phenomena would act to
make populations differ more from each other and from groups less influenced
by these forces. Therefore, why are native Americans so similar to each other
and the Chinese? One possibility is that instead of a single migration wave,
several migration waves from the same source population took place. Also,
recent and generalized migrations from Asia may have helped to keep the
homogeneity of the native American genetic stock and its close genetic af-
finity to the Chinese.

Acknowledgments We dedicate this paper to the memory of Douglas H. Batzer.

Special thanks go to Thomas A. Breslin for his expertise in the history of Chinese
science. We are grateful to all the individuals from the different populations who
contributed to this study. We thank D. Ceballos, M. Ceballos, P. Ioannou, G.M. Troup,
J.R. Kidd, R.M. Fourney, I. Balazs, E. Berminghan, and C. Kolman for samples. The
technical assistance of Oriana Batista in the analysis of the Ngobe population is highly
acknowledged. We thank Frank Verde and Rafael Diaz for their editorial assistance.
This research was supported by the National Institutes of Health through grant
RR08205 awarded to R.J. Herrera and grant ROI HG 00770 awarded to P.L. Deininger
and by the National Science Foundation through grant BNS 90-20567 awarded to M.
Stoneking.

Received 20 September 1996; revision received 6 March 1997.

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