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Post by Admin on May 21, 2020 2:10:12 GMT
The Native Americans of the Pacific Northwest have always claimed to have deep roots in the region. Now, an ancient mariner may be able to back that claim up. Scientists sequencing the DNA of 10,300-year-old human remains from On Your Knees Cave in Alaska have found that he was closely related to three ancient skeletons found along the coast of British Columbia in Canada. These three ancient people were in turn closely related to the Tsimshian, Tlingit, Nisga’a, and Haida tribes living in the region today. The new finding reveals a direct line of descent to these tribes, and it shows—for the first time from ancient DNA—that at least two different groups of people were living in North America more than 10,000 years ago. The study started 21 years ago with an unusually friendly collaboration between archaeologists and the Tlingit tribe that lived near the mariner’s remains on Prince of Wales Island in Alaska. Researchers gathered DNA from the 10,300-year-old skeleton known as Shuká Káa (“Man Ahead of Us”), initially focusing on maternally inherited mitochondrial DNA (mtDNA). They didn’t find a match between its mtDNA and members of the tribe, but they discovered his seafaring ways because isotopes from his teeth showed he ate a marine diet. The project ended on a note of good will between scientists and Native Americans, with a ceremonial reburial of the skeleton in 2008. But as methods of sequencing ancient DNA got exponentially better, the team of geneticists requested permission from the Tlingit and Haida of Alaska, as well as tribes farther south in British Columbia, to extract nuclear DNA from Shuká Káa and three other ancient skeletons. They were allowed to sample the last remaining tissue from Shuká Káa’s molars and from the teeth of a 6075-year-old skeleton on Lucy Island in British Columbia (just 300 kilometers from On Your Knees Cave), a 2500-year-old skeleton from Prince Rupert harbor in British Columbia, and another 1750-year-old skeleton from the same area. Although Shuká Káa’s DNA was too damaged to sequence his entire genome, a team led by molecular anthropologist Ripan Malhi from the University of Illinois in Champaign was able to sequence markers that represented about 6% of his genome. (They also sequenced one-third to two-thirds of the genomes of the other three individuals.) The team then compared those markers to see how closely related the ancient individuals were to each other and to 156 indigenous groups living around the world today. They found that genomewide sequences from the three most recent remains from Lucy Island and Prince Rupert harbor off the coast of British Columbia were closely related to the Tsimshian and other tribes in the Pacific Northwest. Shuká Káa, on the other hand, appears more closely related to groups living in South and Central America today, such as the Karitiana, Suruí, and Ticuna of Brazil’s Amazon. But the signal is not statistically strong, and it may simply be a sign that the tribes all share DNA from the same ancient ancestors in Asia or Beringia where humans lived before they entered the Americas. But Shuká Káa’s maternally inherited mtDNA and nuclear DNA both suggest he is also close kin of the younger skeletons in the study. Connecting the dots between all the ancient individuals, Malhi’s team proposes that Shuká Káa is also ancestral to all those groups, including the Tsimshian and related tribes in the Pacific Northwest, they report today in the Proceedings of the National Academy of Sciences early edition. In another interesting twist, the team found that the group of skeletons was not closely related to two other famous Paleoindians: the 8545-year-old Kennewick Man, unearthed from the banks of the Columbia River in Washington state, and the 12,600-year-old Anzick child from Montana. This suggests there were at least two groups of settlers who came to North America across the Bering Strait land bridge before 10,000 years ago, Malhi says. Although multiple migrations have long been documented, this is the first ancient DNA evidence of different groups arriving in North America at such an early date. Other researchers call the link between the three more recent skeletons and the tribes of the Pacific Northwest “quite clear.” Paleogeneticist Pontus Skoglund of Harvard University, who was not involved in the work, says support for those tribes’ relation to Shuká Káa is weaker, but possible. It could also be, he adds, that Shuká Káa was alive before the lineage formed that led to current northwest groups. Whether the ancient mariner is a direct ancestor of today’s tribes or not, the discovery of the link from the recent skeletons fits with Tlingit and Tsimshian oral traditions that suggest the Tsimshian migrated west along British Columbia’s Naas River to the coast, before spreading north and south, says Rosita Worl, president of the Sealaska Heritage Institute in Juneau and a member of the Tlingit tribe. Archaeologist Timothy Heaton of the University of South Dakota in Vermillion, whose team discovered Shuká Káa, adds that the genetic links fit with the engraving the Tlingit put on the headstone when they reburied Shuká Káa: “We Have Lived in Southeast Alaska Since Time Immemorial. Shuká Káa is Testimony to Our Ancient Occupancy of This Land.” Posted in: ArchaeologyEvolution doi:10.1126/science.aal1002
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Post by Admin on May 23, 2020 5:30:45 GMT
Ancient individuals from the North American Northwest Coast reveal 10,000 years of regional genetic continuity John Lindo, Alessandro Achilli, Ugo A. Perego, David Archer, Cristina Valdiosera, Barbara Petzelt, Joycelynn Mitchell, Rosita Worl, E. James Dixon, Terence E. Fifield, Morten Rasmussen, Eske Willerslev, Jerome S. Cybulski, Brian M. Kemp, Michael DeGiorgio, and Ripan S. Malhi Significance The peopling of the Americas has been examined on the continental level with the aid of SNP arrays, next generation sequencing, and advancements in ancient DNA, all of which have helped elucidate evolutionary histories. Regional paleogenomic studies, however, have received less attention and may reveal a more nuanced demographic history. We present genome-wide sequences of individuals from the northern Northwest Coast covering a timespan of ∼10,000 years and show that continental patterns of demography do not necessarily apply on the regional level. Compared with existing paleogenomic data, we show that geographically linked population samples from the Northwest Coast exhibit an early ancestral lineage and find that population structure existed among Native North American groups as early as the late Pleistocene. Abstract Recent genomic studies of both ancient and modern indigenous people of the Americas have shed light on the demographic processes involved during the first peopling. The Pacific Northwest Coast proves an intriguing focus for these studies because of its association with coastal migration models and genetic ancestral patterns that are difficult to reconcile with modern DNA alone. Here, we report the low-coverage genome sequence of an ancient individual known as “Shuká Káa” (“Man Ahead of Us”) recovered from the On Your Knees Cave (OYKC) in southeastern Alaska (archaeological site 49-PET-408). The human remains date to ∼10,300 calendar (cal) y B.P. We also analyze low-coverage genomes of three more recent individuals from the nearby coast of British Columbia dating from ∼6,075 to 1,750 cal y B.P. From the resulting time series of genetic data, we show that the Pacific Northwest Coast exhibits genetic continuity for at least the past 10,300 cal y B.P. We also infer that population structure existed in the late Pleistocene of North America with Shuká Káa on a different ancestral line compared with other North American individuals from the late Pleistocene or early Holocene (i.e., Anzick-1 and Kennewick Man). Despite regional shifts in mtDNA haplogroups, we conclude from individuals sampled through time that people of the northern Northwest Coast belong to an early genetic lineage that may stem from a late Pleistocene coastal migration into the Americas. ancient DNApaleogenomicsNative Americanindigenouspeopling The initial peopling of the Northwest Coast has received much attention because of its proximity to Beringia and associated implications for an initial coastal migration into the Americas (1⇓–3). Genetic clues for the peopling of the Northwest Coast, however, may be obscured by later demographic events in the region. Studies based on mtDNA and Y-chromosomal markers suggest that populations in the region likely experienced admixture from other groups that entered the region after the initial peopling (4⇓–6). Studies using genome-wide data (7⇓–9) inferred ancient gene flow into North America likely stemming from subsequent movements after the initial settlement. However, because of the limited genomic data from populations in this geographic region, those studies leave questions regarding the degree of temporal genetic continuity of Northwest Coast populations. In the Americas, the oldest thus far whole genome stems from Anzick-1, dating back to ∼12,600 calendar (cal) y B.P. and reportedly associated with Clovis technology (10, 11). Anzick-1 has proven to be surprising in a broader genetic sense, showing greater affinity with Central and South American groups than with Northern groups, despite the ancient burial existing in North America (but comparative indigenous populations from the United States are currently lacking). Shuká Káa, unearthed from On Your Knees Cave (Prince of Wales Island, AK), is not associated with Clovis culture but instead, is associated with a maritime tradition consistent with a coastal migration model and has been dated at ∼10,300 cal y B.P. (3). Shuká Káa exhibited the same mitochondrial haplogroup as Anzick-1 (12), suggesting a link in maternal lineage. Approximately 300 km southeast of the On Your Knees Cave archaeological site is Lucy Island off the coast of British Columbia, Canada. This island is the location of an individual, cataloged as 939, who died ∼6,075 cal y B.P. (13). Individual 939 displays genetic affinity to Pacific Northwest coast groups, such as the Coast Tsimshian (henceforth Tsimshian), that currently live in the same region, but it is difficult to reject 939 as ancestral to both North and South American groups (9). The only other ancient genome from North America is the Ancient One (also known as Kennewick Man), unearthed in the US state of Washington and dating back to ∼8,545 cal y B.P. (14). Kennewick Man also displays surprising results as an early Holocene individual who resided in the Pacific Northwest. His mtDNA belongs to the northern North America limited haplogroup X2a, but his nuclear genome shows affinities with Central and South American populations, similar to patterns observed for Anzick-1. However, a direct ancestry test shows the greatest link to living individuals from the Confederated Tribes of the Colville Reservation, a Native population living in the same geographic region as Kennewick Man (14). On a broader scale, numerous areas of the Americas exhibit patterns consistent with genetic continuity of peoples in the same geographic region over time (9). To test hypotheses related to different demographic scenarios for the peopling of the Northwest Coast, we generated a low-coverage genome (including the complete mitochondrial genome) for Shuká Káa from Alaska (Fig. 1 and Table 1). In addition, we generated two ancient low-coverage genomes, 302 and 443, from Prince Rupert Harbor (PRH), British Columbia (Fig. 1 and Table 1) dating to 2,500 and 1,750 cal y B.P., respectively. Along with previously described genomes from the Americas, we test two hypotheses about the peopling of the Northwest Coast. First, we test whether the people of this geographic region show temporal genetic continuity dating back to at least 10,300 cal y B.P. Second, we test whether the ancestors of the Northwest Coast experienced additional gene flow in the mid-Holocene to further explore the previously observed shift in mtDNA haplogroups on the Northwest Coast (13). Fig. 1 Sampling locations of ancient samples and their associated mtDNA haplogroups Community Engagement It is important to note that the interactions between scientists and indigenous community members associated with this study were and continue to be respectful. Shuká Káa is the indigenous name given to the ancient individual found in On Your Knees Cave on Prince of Wales Island in southeast Alaska. His remains were identified in 1996, the same year in which the Ancient One was unearthed from the banks of the Columbia River near Kennewick, WA. However, unlike the antagonistic relationships that were to develop over the handling of the Ancient One’s remains, T.E.F., a US Forest Service archaeologist, and other researchers engaged with Tlingit- and Haida-speaking communities in Alaska and developed strong working relationships with community members (details are in Datasets S1–S4). With appropriate tribal engagement and discussions, our analyses were conducted on some of the last remaining tissue subsampled from Shuká Káa’s molars for DNA analysis before his repatriation to the Tlingit and reburial in his ancestral land. Farther south, J.S.C. and R.S.M. established a partnership with the Metlakatla and Lax Kw’alaams First Nations in 2007 to aid in the study of the population histories of those communities. They are located in the PRH region of British Columbia. As part of the active partnership, J.S.C. and R.S.M. visit the communities on a regular basis to develop research studies and discuss interpretations of results as well as manuscripts written for peer review publication. The First Nations agreed to allow destructive DNA methods of samples of ancestral individuals analyzed in this study.
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Post by Admin on May 23, 2020 20:10:48 GMT
Results A Mitochondrial Genome Reassessment. The individuals analyzed exhibit DNA damage patterns consistent with ancient DNA (Fig. S1). The complete mitochondrial genome of the Shuká Káa individual belongs to haplogroup D4h3a and was compared with 52 modern (available in GenBank) and 2 ancient D4h3 mitochondrial genomes (939 and Anzick-1) (11, 13). The resulting tree (Fig. S2) clearly shows that Anzick-1 is ancestral to the entire D4h3a clade, whereas the ancient Northwest Coast mitochondrial genomes belong to two different subbranches known as D4h3a9 and D4h3a12, with the latter here defined and encompassing a modern sample of an individual currently living in Bolivia (SI Text). Fig. S1. DNA damage patterns for the ancient individuals. Random subset of all mapped reads for each ancient individual. The mismatch frequency is relative to the reference as a function of read position: C to T in red and G to A in blue.] Fig. S2. Phylogeny of mtDNA haplogroup D4h3. The topology was inferred by maximum parsimony. An ML timescale is indicated below the tree. Insets show the geographic origin of samples, age estimates for the main nodes, and the Bayesian skyline plot. Neff indicates the effective number of females. Ancient mitochondrial genomes are circled in red. Today, the haplogroup D4h3a is virtually absent in northern North America. To the contrary, the mitochondrial genomes of the more recent ancients from the Northwest Coast (443 and 302) are classified as A2 (Table 1), the most commonly reported mitochondrial haplogroup of native North America. Thus, based on the mtDNA data alone, it might be plausible that the native people of the northern Northwest Coast experienced a drastic change in their mtDNA gene pool in a rather short period, possibly because of additional gene flow in the mid-Holocene (mitochondrial genome change hypothesis). However, considering that the mtDNA haplogroup frequencies are likely to change radically because of drift in small populations over time (15), such a mitochondrial genome discontinuity might simply be the result of the limited number of complete mitochondrial genomes analyzed from the area, particularly from ancient individuals (16). Thus, because the mitochondrial genome can describe only part of the ancestral genetic history of the Northwest Coast, we extended the analyses to the entire genome to test alternative hypotheses. Autosomal Genome Assessment. We used outgroup ƒ3 statistics to assess the shared ancestry among the ancient individuals and 169 worldwide populations (9). Outgroup ƒ3 statistics of a worldwide dataset show that all four ancient individuals (Shuká Káa, 939, 443, and 302) display greater affinity with Native American groups than with other worldwide populations (Fig. 2A and Fig. S3). Ranked outgroup ƒ3 statistics suggest that 939, 443, and 302 tend to share greatest affinity with Northwest Coast groups, whereas Shuká Káa ostensibly shows closer affinity to groups farther south (Fig. S4). However, because of the low coverage of the Shuká Káa sample, the relationship is not statistically significant. Fig. 2. Genetic affinity of Shuká Káa and the other Northwest Coast prehistoric humans to global and regional indigenous populations. (A) Heat map represents the outgroup ƒ3 statistics estimating the amount of shared genetic drift between Shuká Káa and each of 156 contemporary populations since their divergence with the African Yoruban population. (B) Maximum likelihood tree generated by TreeMix using whole-genome sequencing data from Raghavan et al. (9) and with the Tsimshian genome masked for European ancestry. (C) Principal components analysis projecting Shuká Káa, 939, 302, 443, Anzick-1 (11), Saqqaq (30), and Kennewick Man (14) onto a set of non-African populations from Raghavan et al. (9), with Native American populations masked for nonnative ancestry. (D) Cluster analysis generated by ADMIXTURE for a set of indigenous populations from the Americas, Siberia, the Arctic, and Greenland and the Anzick-1, Kennewick, MA-1 (19), Saqqaq, Shuká Káa, 939, 302, and 443 samples. The number of displayed clusters is K = 8, which was found to have the best predictive accuracy given the lowest cross-validation index value. Fig. S3. Ranked outgroup ƒ3 statistics examining shared genetic history: (A) 302 and (B) 443 both show a close relationship to indigenous populations from the Northwest Coast, and (C) 939 also has close affinity to individuals from the Northwest Coast but also, the Yaqui from the American Southwest. (D) Shuká Káa shows a close affinity to populations from the American Southwest; however, the rank is not statistically significant. Fig. S4. Genetic affinity of the ancient individuals to contemporary human populations: (A) 302, (B) 443, and (C) 939 all show greater genetic affinity with Native American groups than other global populations. Heat maps summarize the outgroup ƒ3 statistics that estimate the amount of shared genetic drift between each ancient individual and each of 156 contemporary populations since their divergence with the African Yoruban population. To further elucidate the relationship among the ancient individuals of the Northwest Coast and their relationship to modern populations, we examined maximum likelihood trees created with TreeMix (17). C/T and G/A polymorphic sites were removed from the dataset to guard against the most common forms of postmortem DNA damage (18). We observe that 302 and 443 form a sister clade to the modern Tsimshian (masked for European ancestry) (Fig. S5 A and B, respectively). Individual 939 is an outgroup to both North and South Americans (Fig. S5C) as is Shuká Káa (Fig. 3B). However, adding a migration event introduces an edge connecting Europeans and Shuká Káa, which leads to Shuká Káa forming a clade with the Tsimshian and Athabascan (Fig. S6). The signal may represent Native American dual ancestry (19) or be a result of possible contamination (Table S1).
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Post by Admin on May 24, 2020 6:42:58 GMT
Fig. 3. Hypothetical scenarios for the regional peopling of the Northwest Coast. (A) Scenario tested by the D statistic where Anzick-1 is basal to both Shuká Káa and South America, which is rejected, indicating a closer affinity to South America. (B) Scenario tested by the D statistic where Shuká Káa is basal to 939 and both contemporary and ancient Northwest Coast individuals, which is not rejected. (C) Scenario tested by the D statistic where Shuká Káa is basal to ancient and modern Northwest Coast and South America, which is not rejected. (D) Scenario tested by the D statistic where 939, 443, and 302 are basal to the Arctic (Yup’ik and Inuit) and contemporary Northwest Coast populations, which is rejected, indicating a closer affinity to contemporary Northwest Coast populations. Supporting D statistics are listed in Table S2. Fig. S5. ML trees. ML tree generated by TreeMix using whole-genome sequencing data, with the Tsimshian genome masked for European ancestry: (A) 302, (B) 443 and (C) 939. Fig. S6. ML tree with Shuká Káa and one migration. Samples and parameters are the same as in Fig. 2B. Adding the migration event causes Shuká Káa to fall into the Pacific Northwest clade (Athabascan and Tsimshian), with an edge connecting Shuká Káa and Europeans. This result may be a product of contamination (Table S2). Principal components analysis reveals a tight clustering of 939, 443, and 302, which also overlap with modern North American indigenous populations (Fig. 2C and Fig. S7). Shuká Káa falls in close proximity but overlaps with both North and South American groups. The admixture clustering analysis shows a more complicated pattern, where Shuká Káa exhibits mainly components identified in North American and Siberian/Arctic individuals as well as smaller fractions found in South American populations (Fig. 2D). However, individuals 939, 302, and 443 all exhibit a major component found in North American populations.
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Post by Admin on May 24, 2020 21:38:39 GMT
Fig. S7. Principal components (PC) analysis (PC2 and PC3). Shuká Káa, 939, 302, 443, Anzick-1 (11), Saqqaq (30), and Kennewick Man (14) are projected onto a set of non-African populations from the work by Raghavan et al. (9), with Native American populations masked for nonnative ancestry. To further test the hypothesis that people of the Northwest Coast exhibit a close genetic relationship with ancient individuals from the same region, we used the D statistic (20). Given the TreeMix admixture results between Shuká Káa and European populations, we performed a contamination correction to the D statistic as described in the work by Raghavan et al. (19) using observed D statistics with European populations (SI Text). Hypothetical scenarios based on the D statistics are depicted in Fig. 3. The D statistic does not support a scenario of genetic continuity between Anzick-1 and Shuká Káa with respect to South Americans (Fig. 3A and Table S2). The relationship of Shuká Káa, however, is more complex when examined with specific North American ancient and modern groups. Comparing Shuká Káa with 939 and the more recent ancient individuals from the Northwest Coast (443 and 302), we cannot reject an equally diverged relationship with respect to Shuká Káa (Fig. 3B and Table S2). However, we see the same relationship when the comparison is performed between both ancient and modern Northwest Coast individuals and individuals from South America, where Shuká Káa is basal to both groups (Fig. 3C and Table S2). Individual 939 displays a similar pattern with TreeMix and D statistics (Fig. S5C and Table S2), where the individual seems basal to both the Northwest Coast and South America. However, the admixture results show a predominately “North American” component, and contamination-corrected D statistics for 939 indicate a significant relationship toward the Northwest Coast (Table S2, tests 17 and 18). The D statistic did not reveal a signal of gene flow between Arctic populations (Inuit and Yup’ik) and either the modern or ancient Northwest Coast populations compared with Shuká Káa (Table S2, tests 19–28). However, comparing the more recent ancient individuals, the tree was rejected with 939, 302, and 443, indicating greater affinity toward the Northwest Coast populations than the Arctic (Fig. 3D and Table S2). Because certain D (Table S2, tests 8 and 9) and ƒ3 (Fig. S3D) statistics yielded nonsignificant results, we next wanted to examine whether the basal relationship that Shuká Káa exhibited to Northwest Coast and South American populations could be caused by its age relative to the split time of those groups. To address this hypothesis, we simulated genetic data with FastSimCoal2 (21), which allowed us to sample Shuká Káa 10,300 y in the past (SI Text). We considered one scenario (scenario 1), in which Shuká Káa is on the branch leading to the Northwest Coast, and another scenario (scenario 2), in which the sample was on a branch that diverged earlier than the split of the Northwest Coast and South American populations (Fig. S8A). Results for 1,000 simulated replicates under each scenario are plotted in Fig. S8B, indicating that only a small fraction of simulated replicates from scenario 1 could reject the null hypothesis that Shuká Káa is equally diverged to the Northwest Coast and South American lineages, although the simulations placed Shuká Káa on the Northwest Coast branch. However, the reason for this lack of power may be because of the amount of data (which we controlled to yield a similar number of D statistic informative sites as the empirical data). We, therefore, also considered a set of simulations where we increased the expected number of D-statistic informative sites by an order of magnitude. Results from these simulations (Fig. S8C) show that the clear majority of simulated replicates from scenario 1 could reject the null hypothesis that Shuká Káa is equally diverged to the Northwest Coast and South American lineages, with Shuká Káa having higher affinity to the Northwest Coast. Furthermore, results for scenario 2 indicate that the null hypothesis is generally not rejected as expected from the simulated scenario (Fig. S8 B and C). Fig. S8. Simulated Z-score distributions for different scenarios relating Shuká Káa to Northwest Coast and South American lineages. (A) Two scenarios relating Shuká Káa to Northwest Coast and South American linages. Scenario 1 places Shuká Káa on the branch leading to Northwest Coast groups. Scenario 2 places Shuká Káa ancestral to both Northwest Coast and South American groups. (B) Distributions of Z scores under each scenario, in which each distribution is computed from 1,000 independent simulated replicates and where each replicate is based on 200 independent 100-kb genomic regions. The number of D-statistic informative sites is similar to the number in our empirical dataset with Shuká Káa. Only a small fraction of simulated replicates for scenario 1 is able to reject the null hypothesis, although Shuká Káa is on the Northwest Coast branch. In contrast, the majority of replicates for scenario 2 cannot reject the null hypothesis as expected. (C) Distributions of Z scores under each scenario, in which each distribution is computed from 1,000 independent simulated replicates and where each replicate is based on 200 independent 100-kb genomic regions. The number of D statistic informative sites is an order of magnitude larger than in B. A large fraction of simulated replicates for scenario 1 is able to reject the null hypothesis, indicating that Shuká Káa has higher affinity to the Northwest Coast than to the South American lineage. Therefore, more data provided us with the power to reject the null hypothesis. In contrast, the majority of replicates for scenario 2 still cannot reject the null hypothesis as expected. We next considered models that examine a less direct relationship between Shuká Káa and the more recent ancient individuals in addition to the spread of the D4h3a mtDNA haplogroup across the Americas. Because Anzick-1 from Montana predates Shuká Káa (∼12,600 vs. ∼10,300 y B.P.) and shares the same mtDNA haplogroup, we performed D statistics to assess their relationship. The tree with Shuká Káa and 939 as sisters relative to Anzick-1 is not rejected (Fig. S9A). The trees with modern Northwest Coast (mtDNA haplogroup A2) as sister to either 443 (mtDNA haplogroup A2) or 302 (mtDNA haplogroup A2) relative to 939 (mtDNA haplogroup D4h3a) are also not rejected (Fig. S9B). These results are inconsistent with a change in the overall gene pool of the Northwest Coast after the early Holocene, although it led to the observation of very different mitochondrial haplogroups detected over time in the region. We also explored the relationship of Shuká Káa to Kennewick Man (14), who was unearthed in Washington and dates to ∼8,545 cal y B.P. Although Kennewick Man does not share the same mtDNA haplogroup with Shuká Káa, they coexisted within 1,700 y of each other (about 68 generations). In all tests with the D statistic, Shuká Káa displayed a basal relationship to both ancient and modern Northwest Coast populations with respect to Kennewick Man (Fig. S9C and Table S2).
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