New Advances Reconstructing the Y Chromosome Haplotype of Napoléon the First Based on Three of his Living Descendants

This paper describes the findings of the complete reconstruction of the lineage Y chromosome haplotype of the French Emperor Napoléon I. In a previous study (Lucotte et al., 2013) we reconstructed, for more than one hundred Y-STRs (Y–short tandem repeats), the Y-chromosome haplotype of Napoléon I based on data comparing STR allelic values obtained from the DNA of two of his living descendants: Charles Napoléon (C.N.) and Alexandre Colonna Walewski (A.C.W.); in the present study we compare STR allelic values of C.N. and A.C.W. to those of Mike Clovis (M.C.), a living fifth generation descendant of Lucien (one of Napoléon’s brothers). When compared between M.C., C.N. and A.C.W., STR allelic values are identical for a total of 93 STRs; that permits us to propose those values, for which the three living descendants are identical, as expected allelic values of Napoléon I’s Y-chromosome haplotype. For seven STRs, allele values are variable between M.C., C.N. and A.C.W.; we propose for three of them (DYS442, DYS454 and DYS712) expected allelic values, based on data concerning the allele distributions of these STRs in the population.

In a first study (Lucotte et al., 2011) we determined the Y-chromosome non-recombinant part (NRY) haplogroup of Napoléon, based on genomic DNA extracted from two islands of follicular sheats associated with his beard hairs conserved in the Vivant-Denon reliquary (Lucotte, 2010).This haplogroup, established by the study of 10 NRY-SNPs (single nucleotide polymorphisms), is E1b1b1c1*; an "oriental" haplogroup of origin, as shown by the frequency map of M34 in contemporary European populations (Lucotte and Diéterlen, 2014), the antepenultimate SNP of the E1b1b1c1* differentiation.
In this same first study (Lucotte et al., 2011) we studied the buccal smear DNA of Charles Napoléon (C.N.), the living fourth generation of male descent from Jérôme, for the first 37 NRY-STRs (short tandem repeats) of the Family Tree DNA (FT DNA) kit; that permits us to establish a first Y-STR profile of C.N.This profile is highly indicative of the E1b1b1 haplogroup, because of STR allelic values at the discriminant (from Athey, 2006) Y-markers DYS19 (allele 13) and at DYS464.a,.b,.c and .d (alleles 13,14,15 and 16 respectively); moreover allele values (of 13) at DYS19 and at YCaII.a and .b (19 and 22) are the same for Napoléon (N) and for C.N.
In a second study (Lucotte et al., 2013)  We then proposed (Lucotte et al., 2013) a first reconstruction of the Y-chromosome haplotype of Napoléon, based on the expected STR allelic values obtained from the 124 identical STRs between C.N. and A.C.W.

Methods
Mike Clovis (M.C.) is the propositus (Figure 1) for this study.Buccal swab samples for this DNA donor were collected with informed consent.DNA extraction and STRs typing ("upgrade" for 111 genetic markers) were done according to FTDNA recommendations.Because you know exactly how many generations ago the ancestor lived (Figure 1), it is interesting to see how statistics / probabilities compare with reality.In this particular case: Mike > Cyril > Valentine-Louis Clavering > Louis > Lucien > Charles Bonaparte (= Carlo Buonaparte), the probabilities based on the calculations of the time of the most recent common ancestor (the TMRCA) calculations (from Walsh, 2001) are incorrect: comparing the STRs showing only 6 mismatches, it only estimates the probability that Mike Clovis (328303) and Alexandre Colonna Walewski (218983) shared a common ancestor within the last 1 generation = 0.07%, 2 generations = 1.13%, 3 generations = 4.93%, 4 generations = 12.48%, 5 generations = 23.38%,6 generations = 36.19%,7 generations = 49.27%,…;so, about 36 to 50% probability six generations back.Because of this identity, we can reasonably infer that the 93 allelic values of the above genetic markers correspond to those expected for Napoléon I (because they have remained unchanged for 5/6 generations of remote ancestry).

Differential STRs
Table 2 lists and characterizes the seven STRs that differentiate between M.C., C.N., and A.C.W.Only one of them (CDY.a) is palindromic.The mutation rates, when known (Burgarella & Navascués, 2011), of these differential alleles are in the 10 -3 range (except for DYS447).These rates are impossible to evaluate for the palindromic STR CDY.a, and unknown for the moment for DYS712.We already know (Lucotte & Bouin-Wilkinson, 2014) the real allele value = 23 of Y-GATA-C4 for Napoléon I.

Discussion
In the goal to establish the Y-chromosome haplotype of Napoléon the First we determined initially, in his genomic DNA extracted from two islands of follicular sheats associated with his beard hairs conserved in the Vivant-Denon reliquary (Lucotte et al., 2011) Because of the identity of allelic values of STRs between Charles Napoléon (the living fourth generation of male descent from Jérôme (Napoléon I's youngest brother) and Alexandre Colonna Walewski (a direct living sixth generation descendant from Napoléon I), we proposed (Lucotte et al., 2013) expected allelic values of Napoléon I for a total number of 109 STRs (33 of them being palindromics).For some of the six variables (between Charles and Alexandre) STRs: DYS454, DYS481, Y-GATA-C4, DYS712, CDY.a (palindromic) and DYF397.2(palindromic), we proposed as expected allelic values for Napoléon I the most probable allelic forms according to STR distributions; the allele value of DYS454 = 7 for Charles Napoléon appeared then as a highly discordant one.
Mike Clovis is a living, previously unknown, fifth descendant of Lucien (another brother of Napoléon I).The objective of the present study is to compare, for a total number of 106 STRs, allelic values between him, Charles Napoléon and Alexandre Colonna Walewski.Identity of allelic values between the three was confirmed for 82 non-palindromic STRs and for 11 palindromic STRs; that confirms, in a triangular form, that these 93 STR allelic values are definitely those previously proposed as expected allele values of the Napoléon I Y-haplotype.
These comparisons between Mike Clovis, Charles Napoléon and Alexandre Colonna Walewsky permit us to clarify some of the questions asked by the variable values between them: for DYS454, the allele value = 11 for Mike Clovis is the expected allelic value of Napoléon I, as previously proposed.For DYS712, the allele value = 23 for Mike Clovis corresponds also to the expected allelic value of Napoléon I already proposed; however in this case it is not the modal class of distribution of DYS71 values that is concerned, but the nearest one of the right edge of this distribution.
Compared to Charles Napoléon and Alexandre Colonna Walewski, Mike Clovis had different allele values for DYS442 = 11 and DYS447 = 22.For DYS442, as proposed previously, allele value = 12 is probably the expected allelic value of Napoléon I because it corresponds to the modal class of the distribution; and allele value = 11 for Mike Clovis results from a single mutational event (one-step, minus 1).
It is impossible to predict some expected allelic value of Napoléon I for DYS447, because the three obtained allele values (that of Mike Clovis = 22 could be the result of a one-step plus 1 mutational event) are all located at the left tail of the distribution.It is impossible also to predict some expected allelic value of Napoléon I for DYS481, even when interpreted in the context of the oriental origin of the E1b1b1c1 haplogroup (Lucotte and Diéterlen, 2014), because all the three obtained allele values are now located at the right tail of the distribution.
We ignore, for the moment, what is the Y-chromosome haplotype of Carlo Buonaparte; but it seems highly probable, because of the similarities between the Y-STR values presently obtained, that he was the biological father of Lucien, Napoléon and Jérôme (all these three having the same Y-haplogroup).As a by-product of such studies, we established that the allele value = 7 for DYS545 is highly characteristic of the Jérôme line; possibly, as shown here, the allele value = 11 for DYS442 could be characteristic of the Lucien line.It remains a possibility that allele values of 25 for DYS712 and of 28 for DYS481 could be characteristics of the direct Napoléon I line, at least for the Walewski descent.
we established a more complete (because based on the FTDNA-111STRs kit) Y-STR profile of C.N., and the 111-Y-STRs profile of Alexandre Colonna Walewski (A.C.W.), the fifth generation descendant of Alexandre Walewski (1810-1868) who was the son born of the union between Napoléon I and Countess Maria Walewska (1786-1817).Comparisons at the time between the two STRs profiles were realized for a total number of 130 STRs, six of them (DYS454; DYS481; DYS635 = Y-GATA-C4; DYS712; DYS724 = CDY.aand DYF397.2) having different allelic values between C.N. and A.C.W.At that time we only had three direct determinations available on real allele values of Napoléon (for DYS19 = 13, and for the palindromic YCAII.a=19 and .b= 22).
sixteen supplementary allelic direct determinations (on a lock of hair dandruff dating from 1811) for Napoléon I STRs, in order; DYS 19 = 13; palindromic DYS385.a and b. = 16; DYS389.i= 14, .ii= 31; DYS390 = 24; DYS391 = 10; DYS392 = 11; DYS393 = 14; DYS438 = 10; DYS439 = Y-GATA-A4 = 12; DYS448 = 20; DYS456 = 15; DYS458 = 16; Y-GATA-C4 = 23 and Y-GATA-H4 = 11.These results confirm our previous ones for allele 13 at DYS19; moreover all these other direct determinations (except for Y-GATA-C4) are in accordance with our previous direct predictions (Lucotte et al., 2013) concerning the expected values for the corresponding STRs.Mike Clovis (M.C.) is the fifth generation descendant (Figure 1) of Lucien; to visualize the generations of the two male descent from the Walewski and the Jérôme lines, see the first figure of the Lucotte et al., 2013 article.In order to realize a triangular comparison between three living males related to Napoléon I (a direct descendant: A.C.W.; an indirect descendant from his brother Jérôme: C.N.; and M.C., an indirect descendant from his brother Lucien), we study now in the present article the Y-STR profile of M.C. by means of the FT-DNA -111 STRs kit; and we compare this STR profile to those of C.N. and A.C.W.

Figure 1 .
Figure 1.Chain of transmission (seven successive generations of paternal ancestry) from the ancestor Charles Bonaparte (Napoléon's father) to the propositus (arrow) Mike Clovis

Figure 2
shows the bimodal distribution of Y-GATA-C4 alleles in the population; Napoléon I (N) value corresponds to that of the second modal class.Allele values (=22) of Charles Napoléon (C.N.) and Mike Clovis (M.C.) can be explained admitting one-step (minus 1) mutations, and that (=21) of Alexandre Colonna Walewsky (A.C.W.) admitting a two-step (minus 2) mutation.

Figure 5 .
Figure 5. DYS442 and DYS447 allelic distributions, based on our sample of 1 000 unrelated European Caucasians

Figure 7
Figure 7 shows the modal distributions -based on our sample of 1000 European subjects -of allelic classes for the palindromic markers CDY.a and CDY.b.It is because of the identity of allele values = 36 between C.N., A.C.W.

Figure 7 .
Figure 7. CDY.a and .ballelic distributions, based on the sample of 1 000 unrelated European Caucasians , allelic values for the three Y-STRs DYS19 and for two palindromic STRs YcaII.a and .b. Subsequently (Lucotte and Bouin-Wilkinson, 2014), based on genomic DNA of his hair dandruff dating from 1811, we determined allelic values for 16 STRs: DYS19 (for which we confirmed the first allelic value previously obtained), the palindromic STRs DYS385.a and .b,DYS389.i and ii, DYS390, DYS391, DYS392, DYS393, DYS438, the variable STR-Y-GATA-A4, DYS448, DYS456, DYS458, Y-GATA-C4 and Y-GATA-H4.The corresponding allele values for these 18 STRs correspond to the real allelic values of the Napoléon I Y-haplotype.