DYS464 extended test aka DYS464x plus some comments on DYF399

 

John McEwan

4th June 2006

 

Background

DYS464 is a multi-copy Y chromosome STR, with typically 4 copies, but ranging from 0 to 7, and is offered by several companies in particular FTDNA. The marker is quite difficult to handle from an analysis perspective, because like other multi-copy repeats the standard tests cannot distinguish the individual copies. Convention lists them from lowest to highest. This makes comparison between diverged individuals fraught because it is often not clear which copy has altered. 

 

These tests are also subject to additional issues, because unlike normal markers peak height has to be used to score whether they have one or more copies of a particular allele. Certain combinations of alleles make this difficult, especially if copy number also varies as well eg AABB and ABnullnull.

 

However, in practice the marker has been shown to be extremely useful from a genealogical perspective, both in distinguishing families, and for predicting haplogroup.

 

More recently, an extended test has been developed DNA-Fingerprint that can in some circumstances distinguish between some of the alleles. This is possible because there has been a G->C SNP mutation identified, which “tags” the alleles where it is present. However, this test also identifies a further process that affects variability in this marker called gene conversion or recombinatorial loss of heterozygosity or RecLOH for short. This happens in part because sections of the Y chromosome are arranged and duplicated in palindromes aka inverted repeats so they can repair or delete themselves even though only one copy of the DNA is available. The genomic organization of DYS464 and DYF399 in the AZFc region is shown in Figure 1 below taken from Henson (2005). 

 

 

The mutation rate for SNPs is about once every 50,000,000 generations and for STRs about once every 500 generations: RecLOH events have an intermediate frequency between these events. The paper by Henson also summarizes that these RecLOH events and deletions often affect multiple markers and in this case both DYF399 and DYS464.

 

Variability of DYS464x in the R haplogroup

This section expands and updates a Genealogy DNA listserver message by Gareth Henson in November 2005. Most non R1b1c individuals have 4 DYS464 alleles and the SNP is G i.e. GGGG. At some point a G->C mutation occurred and through RecLOH it was converted to CCGG and then a further duplication to CCCG. The CCCG form is the one most commonly observed in R1b1c individuals. Thus an initial mutation and then two further steps were required. This is almost certainly a unique event. However, the RecLOH process means that additional variability is observed. This makes the test a “quazi-SNP” i.e. less variable than an STR, but more variable than a SNP and it has value both for exploring deep ancestry and as a genealogical marker.

 

Exactly when the DYS464 GGGG form changed to CCCG form is not known, but public results from Ymatch show the GGGG form for haplogroup A (n=1), E (2), G (3), J (1 GGGG, 1 GGnullnull), I (4), R1a (1). The majority of the R1b1c records are of the CCCG form. There is also one R1b1* (P25+, M269-) that has the GGGG form (grey see table 1 below). A M269+, S21-, S28- individual with the GGGG form has also been identified (see R1b SNP table). This suggests that perhaps this change most likely happened soon after the M269+ mutation, and before the S21 and S28 mutations as it is unlikely to have had 2 RecLOH events to revert to the GGGG form. However, this hypothesis requires further results to be confirmed.

 

Deletion events in DYS464 may also affect adjacent markers as well especially DYF399S1, DYF371, DYF385, DYF399 and DYF408. Table 1 is an edited extract of R1b from Ymatch with only the multiple copy markers retained that have been scored for DYS464x.  Although care is needed as some entries may be mis-scored (highlighted pink, all with double peaks which are hard to reconcile) it is generally observed that when only two alleles are DYS464 are present: only one allele of DYS724, two alleles of DYS725, three alleles of DYF371, one allele of DYF401 and 3 alleles of DYF408 are also present. Note that CCnullnull (n=3) and CGnullnull (n=2) seem equally prevalent out of 80 R1b DYS464x records available. They also identify that these deletions have happened multiple times. Insufficient records have been collected as yet to identify how diagnostic these markers are, but any R1b DYS464 results that are of the form XXXX are prime candidates for deletion testing and at least DYS464x, DYF399 and DYS371 should be tested together to validate the results. There is equivocal evidence that some of these myriad of partial deletions may reduce male fertility in a minority of cases, but it is clear that deletion of the whole region results in infertility (see Vogt, 2005 for a review and Carvalho et al 2006 for a more recent reference).

 

The other common event is a CCCG -> CCGG or CCCG -> CCCC RecLOH with 7 out of 80 and 1 out of 80 respectively. These events have been observed separately in S21+ and S21- individuals (see R1b SNP table) again suggesting multiple events.

 

In summary, the DYS464x test and the associated DYF399 test provide valuable information especially within R1b and are identifying useful variants for reconciling close matches and also potentially for deeper ancestry.

 

 

Table 1. Ymatch R1b records for multiple copy markers highlighted examples of probable RecLOH events and deletions.

 

References

Carvalho CM, Zuccherato LW, Bastos-Rodrigues L, Santos FR, Pena SD. 2006. No association found between gr/gr deletions and infertility in Brazilian males.

Mol Hum Reprod. 12:269-73

 

DNA-Fingerprint explanation of RecLOH

 

DNA-Fingerprint sample results from DYS464x

 

DNA-Fingerprint Ymatch

 

Henson, G.L. 2005 DYF399S1: A Unique Three-Copy Short Tandem Repeat on the Human Y Chromosome. Journal of Genetic Genealogy 1:8-11

 

Vogt PH. 2005. AZF deletions and Y chromosomal haplogroups: history and update based on sequence. Hum Reprod Update. 11(4):319-36

 

Wikipedia RecLOH

 

Wikipedia repair of the Y chromosome