James A. Spencer

Genes on the short arm of the human X chromosome are not shared with the marsupial X

Eight genes located on the short arm of the human X chromosome (MAOA, SYN1, OAT, OTC, CYBB, DMD, ZFX, POLA) have been mapped in several marsupial species by cell hybrid analysis and/or in situ hybridization using probes derived from human cDNA. Seven appear to be autosomal in all marsupial species examined. The eighth, CYBB, detected a site on the X, as well as major autosomal sites. Although these genes are not conserved on the X chromosome in marsupials, at least some of them are arranged together in autosomal clusters. The autosomal location of human Xp genes in marsupials could mean that this region either was lost from a large ancestral X chromosome in the marsupial lineage or was acquired by a small ancestral X (and perhaps Y) in the eutherian lineage. Either explanation demands that the region was not subject to X chromosome inactivation in a common ancestor 120-150 MyrBP.

Autosomal localization of the amelogenin gene in monotremes and marsupials: implications for mammalian sex chromosome evolution.

We have determined by Southern blot analysis that DNA sequences homologous to the AMG gene probe are present in the genomes of both marsupial and monotreme mammals, although adult monotremes lack teeth. In situ hybridization and Southern analysis of cell hybrids demonstrate that AMG homologues are located on autosomes. In the Tammar Wallaby, AMG homologues are located on chromosomes 5q and 1q and in the Platypus, on chromosomes 1 and 2. The autosomal location of the AMG homologues provides additional support for the hypothesis that an autosomal region equivalent to the human Xp was translocated to the X chromosome in the Eutheria after the divergence of the marsupials 150 million years ago. The region containing the AMG gene is therefore likely to have been added 80-150 million years ago to a pseudoautosomal region shared by the ancestral eutherian X and Y chromosome; the X and Y alleles must have begun diverging after this date.

The X chromosome of marsupials shares a highly conserved region with eutherians

Ten genes, located on the long arm of the human X chromosome, were mapped in several marsupial species by somatic cell analysis and in situ hybridization. All were located on the X chromosome in each species. We conclude that the long arm of the human X chromosome represents a highly conserved region that formed part of the X chromosome in a therian ancestor 120-150 million years ago, before the mammalian infraclasses diverged.

Phosphoglycerate kinase pseudogenes in the tammar wallaby and other macropodid marsupials

Phosphoglycerate kinase (EC 2.7.2.3; PGK) exists in two forms in marsupials. PGK1 is an X-linked house-keeping enzyme, and PGK2 is a mainly testis-specific enzyme under autosomal control. We have used PGK1 probes derived from two closely related species of macropodid marsupials (kangaroos and wallabies) to demonstrate the existence of a large family of pseudogenes in the tammar wallaby (Macropus eugenii). Over 30 fragments are detectable after Taq digestion. We estimate that there are 25–30 copies per genome. Most are autosomally inherited and are apparently not closely linked. Only two restriction fragments that appeared to be sex linked could be detected. Varying degrees of hybridization of fragments to the probes suggest different levels of homology, and hence different ages of origin. The existence of two PGK1 homologous restriction fragments from the X and a large number from the autosomes was also demonstrated by somatic cell hybridization for two other macropodid species, the wallaroo (M. robustus) and the red kangaroo (M. rufus). These results are compared with those from human and mouse, and it is suggested that the propensity of PGK1 to form pseudogenes is an ancient (∼130 MYR BP) characteristic of mammals. The high level of polymorphism detected in the tammar makes these PGK1 probes potentially useful for measuring genetic variability in this species and other macropodids.