Research Article
Austin J Genet Genomic Res. 2015; 2(2): 1015.
Towards a Full-Length Y-Chromosome DNA Sequence of Napoléon the First: Beyond the E-M34 SNP Sub- Haplogroup
Gérard Lucotte¹*, Peter Hrechdakian² and Denis Savard³
¹Institute of Molecular Anthropology, Paris, France
²Unifert Group S.A, Belgium
³469 Chemin Thériault / Bertrand, N.-B. E1W1K5, Canada
*Corresponding author: Gérard Lucotte, Institute of Molecular Anthropology, 44 Monge Street, 75 005 Paris, France
Received: December 22, 2015; Accepted: December 28, 2015; Published: December 31, 2015
Abstract
We report results obtained for the Y-chromosome DNA sequence of Mike Clovis (M.C.), a living fifth generation descendant of Lucien who was one of Napoléon the First (1769-1821)’s brothers. We have previously shown that the Y-haplogroup of Napoléon is E1b1b2a1, SNP (Single Nucleotide Polymorphism) M34 being the terminal genetic marker of this differentiation (Y sub-haplogroup E-M3, in abbreviate). Results obtained here show that there are six successive SNPs (Z841, L791, Y4971, Y4976, Y4970 and PH3893) under M34 in the Y-DNA sequence of M.C. Dating of the corresponding sub-haplogroups shows that E-PH3893 had a TMRCA (Time of the Most Common Recent Ancestor) of about 2700 ybp (years before present). This study permits a better understanding of the paternal ancestry of Napoléon in historical times.
Keywords: Napoléon the First paternal ancestry; Y-chromosome DNA sequence; Single nucleotide polymorphisms; Y-chromosome sub-haplogroups; Time of the most common recent ancestor
Introduction
Mike Clovis (M.C.), is a living fifth generation descendant (Figure 1) of Lucien, one of Napoléon Bonaparte’s brothers. As part of Napoléon I Genome (NIG) project we have sequenced the DNA of the Y-chromosome of M.C., in order to obtain pertinent information about Napoléon’s Y-chromosome.
Our work on Napoléon Y-DNA markers had begun [1] with the obtaining of genomic DNA from authentic Napoléon’s hairs conserved in the Vivant Denon reliquary [2]. Ten successive SNPs (Single Nucleotide Polymorphisms) of the Y-chromosome (M125, M174, M33, M35, M78, M81, M123, M34, M84 and M290) were used to determine Napoléon’s Y-haplogroup, that is E1b1b1c1 according to the human Y-chromosomal haplogroup tree [3]; further studies showed that the SNP M34 is the terminal genetic marker of this differentiation, so [4] the corresponding Napoléon Y-chromosome sub-haplogroup was at that time E1b1b1b2a1 (E-M34 in abbreviate).
In this initial study [1] on Napoléon Y-chromosome, only three Y-STRs (Short Tandem Repeats) were used: DYS19 and YCAII.a and .b (palindromics), the respective allelic values obtained being = 13, 19 and 22. Computed with the haplogroup predictor program [5], these three values considered together estimate that Napoléon I corresponds (with a probable value of 76.5%) to a subject belonging to the previously described Y-haplogroup cluster E1b1b [6]. In that study [1] we determine also allelic values for 37 Y-STRs from buccal smear genomic DNA extracted from Charles Napoléon, the living 4th generation descendant of Jérôme Bonaparte (Figure 1), Napoléon I’s youngest brother. Results showed the allelic values for DYS19 and for YCAII.a and .b are the same for Charles Napoléon and for Napoléon I. Computed with [5], Charles Napoléon’s Y-STR profile is very highly indicative (99.9%) of the E1b1b1c1 Y-haplogroup ; as Charles Napoléon is M34+ , his Y-sub-haplogroup is also E-M34.
In our second study on Y-polymorphisms in Napoléon I’s family [7] we have compared, on a total set of one-hundred and thirty three different Y-STRs, the Y-STR profile of Charles Napoléon to that of Alexandre Colonna Walewsky (who is also E-M34), the living 5th generation direct descendant of Napoléon I (Figure 1). Four nonpalindromic and two palindromic STRs have different allelic values between Alexandre Colonna Walewsky for 106 Y-STRs. In our third study [8] comparing Mike Clovis (also E-M34), Charles Napoléon and Alexandre Colonna Walewsky for 106 Y-STRs, we found variable allelic values between these three subjects for 7 STRs only: for DYS442, DYS447, DYS454, DYS481, DYS635, DYS712 and CDY. a (palindromic). All the observed identical STRs allelic values between the three living subjects, were used to predict the corresponding deduced allelic values for Napoléon I’s STRs. A direct determination of allelic values for 16 STRs of Napoléon [9], determinations that were done on Napoléon I genomic DNA extracted from dandruffs adherent to a Napoléon’s lock of hair dating from the year 1811, allow us to obtain the real allelic values of Napoléon’s STRs for 15 supplementary STRs (allelic value at DYS19 confirmed) others than the three found initially [1].
Figure 1: Chain of transmission (seven successive generations of paternal
ancestry) from the ancestor Charles-Marie Buonaparte (Napoléon’s father)
to the propositus (arrow) Mike Clovis (M.C.). Are also indicated in the tree
the successive generations of paternal ancestry for Alexandre Colonna-
Walewski and for Charles Napoléon.
We have constructed an isofrequency map of the Y-SNP M34 [10], using a large set of genomic DNAs of unrelated males originating from various countries in Europe, Northern Africa and the Near East. The genetic landscape obtained is that of a complex pattern of clines and local decreases and increases in M34 frequencies in some areas. In Armenia, the M34 percentage is 4.2%. The maximal peak in frequency (31%) observed in this map, located in the Dead Sea region of Jordany, indicates the location of origin of M34 in the corresponding region of the Levant. We have reconstructed past migratory routes of M34 chromosomes from this center of origin to West Europe via: Turkey (6.9% in Southeastern Turkey; 7.8% in The Istanbul region), Greece (3.8% for Athens; 1.8% for Northern Greece), the Balkans (1.6% for Croatia) and the continental part of Italy (1.5%).
A secondary peak (9 on 111 subjects = 8.1%) of M34 frequencies, in Ajaccio (Napoléon’s native town), can be observed in the partial isofrequency map we have constructed for Corsica and the surrounding regions; the Calabrian region, located in the south of continental Italy, had the most important M34 frequency (5.5%) after that of Ajaccio.
We know 18 generations of Napoléon’s paternal ancestor’s [11]. His remote ancestor, Gianfardo, was born and lived in Sarzana (a small Italian town on the south of La Magra, a river separating Liguria and Toscany) between the end of XIIth and the beginning of the XIIIth Century. Giovanni (11th generation) was the first paternal ancestor of Napoléon to leave Sarzana for Corsica. None of the nine Ajaccian subjects (and not the only Bastian subject) we have studied -bearing the M34 marker- has the Bonaparte’s patronym.
Material and Methods
Buccal swab samples for M.C. were collected, with informed consent. His Y-chromosome sequence was studied in the framework of the “Big Y” project [12], conducted by the Family Tree DNA (FTDNA) Company.
Sequencing was performed on the Illumina HiSeq platform, and downstream analysis was conducted with Arpeggi genome analysis technology. After the sequencing procedure the reads were mapped to the Hg19 version of human genome reference [13], followed by post-processing and variant calling ; all of which were performed using a software (Arpeggi engine).
Aligned data concerning M.C. are given in the form of a BAM file (https://www.dropbox.com/s/nir3pqf6073gdnr/4464.bam.zip?dl=0). This BAM file (after YFull treatment) was downloaded from the site of one of us (D.S.), under the reference E-PH3893.
The M.C. Y-chromosome samples were sequenced at an average depth of 57.76x (Table 1). Centered on the male specific region of the Y-chromosome (MSY), the sequenced part corresponds approximately to 55.25% of the total of the Y-chromosome sequence.
BAM file size
01Gb
Read numbers
7601387
Mapped reads
All
Length coverage
14173443 bp
Minimal and maximal depth coverage
1x-7999x
Mean depth coverage
57.76x
Median depth coverage
39x
Table 1: Main Characteristics of the M.C.’s Y-chromosome sequence obtained.
Results
A mean of about 30 000 SNPs are studied in this sort of Big Y analysis [14]. At first time, we made sure that M.C. is really M34+ (Table 2).
Sub-haplogroups
SNPs found
E-M34
M34 + Y2931, Y2929, Y4143…+ 57 other SNPs
E-Z841
Z841 + Y2961, CTS2339
E-L791
L791 + L792, Z838, Z852… + 86 other SNPs
E-Y4971
Y4971 + Y4973, Y4968, Y4975
E-Y4976
Y4976 + Y4972, K257
E-Y4970
Y4970
E-PH3893
PH3893+PH4283, Y12966, Y12962… + 9 other SNPs
Table 2: Successive M.C. sub-haplogroups below E-M34, and Y-SNPs found on it.
A total number of 491 STRs were studied in the present study. The 93 STR allelic values of M.C. found were identical to those published [8]. Particularly, the variable M.C. allelic values DYS442=11, DYS454=11, DYS481=27, DYS635=22, DYS712=23, and CDY.a = 34 are the same. This establishes that it is well the Y-chromosome sequence of M.C. that is under study.
Table 2 summarizes the lists of Y-SNPs of the M.C. Y-chromosome sequence found in the same drafts corresponding to (depicted on Figure 2) the six successive Y-sub-haplogroups (E-Z841, E-L791, E-Y4971, E-Y4976, E-Y4970 and E-PH3893) located below E-M34. In these drafts the sub-haplogroup E-Y4970 is characterized by the corresponding SNP only, while there is a set of two supplementary SNPs for E-Z841 (Y2961 and CTS2339) and for E-Y4976 (Y4972 and K257), a set of three (Y4973, Y4968 and Y4975) for E-Y4971, and up to 89 (L792, Z838, Z852 and 86 others) for E-L791.
The last found in this serie (the most recent, see Table 3) of M.C. subhaplogroups is E-PH3893, with PH4283, Y12966, Y-12962 and nine other SNPs.
Sub-haplogroups below E-M34
Datations
E-Z841
Formed 14 900 ybp ; TMRCA = 14 000 ybp
E-L791
Formed 14 000 ybp ; TMRCA = 5 600 ybp
E-Y4971
Formed 5 600 ybp ; TMRCA = 5 300 ybp
E-Y4976
Formed 5 300 ybp ; TMRCA = 4 800 ybp
E-Y4970
Formed 4 800 ybp ; TMRCA = 4 700 ybp
E-PH3893
Formed 4 700 ybp ; TMRCA = 2 700 ybp
Table 3: Dating estimates of the six sub-haplogroups below E-M34.
Figure 2: Schematization of the successive Y-SNPs under M34 found in the
Y-DNA sequence of Mike Clovis. M. Clovis, Shirokoff and Gre-17 belong to
the E-PH3893 Y sub-haplogroup; Savard and del Turco belong to E-Y4970.
Figure 2 is a schematization, according to [15], of the filiation –relationships between the six successive M.C. SNPs located under M34. M.C., belonging to sub-haplogroup E-PH3893, is characterized by S7897 and seventeen of other SNPs.
Up to now there are six subjects, belonging to the latest E-PH893 and E-Y4970 sub-haplotypes, that are involved in the Big Y procedure [16]. M. Clovis, and Shirokoff (of Russian origin), are two of the three subjects constituting the E-PH3893 group (Figure 2). Shirokoff is characterized by F3099, PH460, PH1181, CTS913, FGC4198 and by twenty-six other YFS-SNPs.
Previous estimates concerning E-M34 datation in Armenia [17] are up to circa 4.8 ±0.8 to 5.1±1.2 kya. Table 3 gives estimates, among [18], concerning dating in ybp (years before present) and TMRCA (Times of the Most Common Recent Ancestor) for the seven M.C. sub-haplogroups below E-M34. Based on the number of SNPs common to M. Clovis and Shirokoff: PH3893, PH4289, Y12961 to Y12969, Y13512 to Y13514, PF498rc (reccurent), M4081rc and Y1829rc, the TMRCA is about 2 700 ybp.
The third individual belonging to sub-haplogroup E-PH3893 studied in the Big Y project is gre-17 (originating from Greece), that is characterized by F3099 and thirty other SNPs (Figure 2).
One of us (D.S.), whose sub haplogroup derives from E-Y4970 (and consequently is the nearest neighbour of the three previous subjects belonging to E-PH3893 sub-haplogroup found in the Big Y project), is of distant Armenian origin. He is characterized by L884, L128, FGC1551, PF3499, Z54459, M7761 and twenty- nine other FS SNPs. Also deriving from E-Y4970 is del Turco (M2783-), whose name suggests an ancient Turkish origin.
Conclusion
One of our studies [1], based from genomic DNA extracted from his remains, permits us to obtain the Y-chromosome haplogroup of Napoléon the First. This Y-haplogroup is E1b1b2a1, according to present–day rules of Y-haplogroup nomenclature. As the terminal SNP marker of the differentiation is M34, we designated this corresponding Y sub-haplogroup (in abbreviate form) as E-M34. Based on a large number of unrelated DNA of subjects originating from Europe, Northern Africa and the Near East [10], we constructed a global isofrequency map of the M34 genetic marker. The geographic origin of M34 is some region in the Levant. The past migratory routes of M34 chromosomes from this center of origin to West Europe are Turkey, Greece, the Balkans and the continental part of Italy; that links with the surrounding regions of Corsica, in Ajaccio, where Napoléon was born. Early estimates of E-M34 datation [17] concerned a period approximately in the middle of the Neolithic.
Mike Clovis (M.C.) is a living fifth generation descendant of Lucien, one of Napoléon’s brother. He is E-M34, and his Y-STR pro le is quasi-similar to those of the direct descendant of Napoléon or of a Napoléon’s brother [1,7,8], as well as that of Napoléon’s deduced profile. In the present study the full-length Y-chromosome DNA sequence of M.C. was determined by the big Y procedure. Six successive SNPs under M34 were detected in the M.C. Y-chromosome DNA sequence. The corresponding E sub-haplogroups were dated, according to the Time of the Most Common Recent Ancestor (TMRCA) methodology ; this dating evolve from 14000 years before present (ybp) for E-Z841 to 4700 ybp for the antepenultian E-Y4970 sub-haplogroup.
The estimated TMRCA of the most recently found M.C. subhaplogroup E-PH3893 is about 2700 ybp. That time interval concerns the historical time when Napoléon male ancestors evolved. It seems remarkable that several subjects included in this big Y study belong to the various countries located among the migratory road starting from the M34 focus in the Levant [10] that finally goes to Corsica.
Acknowledgment
We thank Mike Clovis, who furnished to us his personal BAM file. The sequencing work was realized by the Family Tree DNA Company, in the frame of the Big Y project. We thank the Count Alexandre Colonna Walewski for his indefectible financial support.
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