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Background  Maleness in mammals is genetically determined by the Y chromosome. On the Y chromosome  SRY  is known as the mammalian male-determining gene. Both placental mammals (Eutheria) and marsupial mammals (Metatheria) have  SRY  genes .  However, only eutherian  SRY  genes have been empirically examined by functional analyses, and the involvement of marsupial  SRY  in male gonad development remains speculative.    Results  In order to demonstrate that the marsupial  SRY  gene is similar to the eutherian  SRY  gene in function, we first examined the sequence differences between marsupial and eutherian  SRY  genes. Then, using a parsimony method, we identify 7 marsupial-specific ancestral substitutions, 13 eutherian-specific ancestral substitutions, and 4 substitutions that occurred at the stem lineage of therian  SRY  genes. A literature search and molecular dynamics computational simulations support that the lineage-specific ancestral substitutions might be involved with the functional differentiation between marsupial and eutherian  SRY  genes. To address the function of the marsupial  SRY  gene in male determination, we performed luciferase assays on the testis enhancer of Sox9 core (TESCO) using the marsupial  SRY . The functional assay shows that marsupial  SRY  gene can weakly up-regulate the luciferase expression via TESCO.    Conclusions  Despite the sequence differences between the marsupial and eutherian  SRY  genes, our functional assay indicates that the marsupial  SRY  gene regulates  SOX9  as a transcription factor in a similar way to the eutherian  SRY  gene. Our results suggest that  SRY  genes obtained the function of male determination in the common ancestor of Theria (placental mammals and marsupials). This suggests that the marsupial  SRY  gene has a function in male determination, but additional experiments are needed to be conclusive.    Electronic supplementary material  The online version of this article (10.1186/s12862-018-1119-z) contains supplementary material, which is available to authorized users.

The evolutionary process of mammalian sex determination genes focusing on marsupial SRYs