title: Gene structure and identification of minimal promoter of Pou2 expressed in spermatogonial cells of rohu carp, Labeo rohita.
abstract: Mammalian Pou5f1 is a known transcriptional regulator involving maintenance of embryonic and spermatogonial stem cells. Little is known about teleost Pou2, an ortholog of mammalian Pou5f1. Evidences of discrepancy in expression pattern between fish species were documented. To better understand, we have cloned and characterized Pou2 gene of farmed rohu carp, Labeo rohita. It contained five exons with an open reading frame of 1419 bp long, translatable to 472 aa. A bipartite DNA binding domain termed POU domain, comprising of POU-specific and POU-homeo sub-domains, was identified. Rohu Pou2 is highly conserved with zebrafish counterpart, as evidenced by 92% overall sequence identity of deduced protein. The POU domain remained highly conserved (showing more than 90% identities) within fish species. Even though there is a divergence between Pou2 and Pou5f1, the common POU-specific domain remained conserved throughout eukaryotes indicating their possible involvements in common trans-activation pathway(s) between mammals and non-mammals. In support, we have provided evidence that Pou2 is indeed abundantly expressed in proliferating rohu spermatogonial cells and hence participates in stem cell maintenance. Its mRNA accumulation in the ovary supported about its maternal transmission with possible regulatory roles during embryogenesis. The 5'-flanking region (~2.7 kb) of rohu Pou2 was sequenced and computational analysis detected several putative regulatory elements. These elements have been conserved among fish species analysed. Luciferase assay identified a mammalian-type 'TATA-less promoter' capable of driving Pou2 gene transcription. These findings will help for future studies in elucidating participatory role of fish Pou2 in male germ cell development.
authors.affiliation: Fish Genetics and Biotechnology Division, Central Institute of Freshwater Aquaculture, Indian Council of Agricultural Research, Kausalyaganga, Bhubaneswar, 751002, Odisha, India.
authors.forename: Chinmayee
authors.initials: C
authors.lastname: Mohapatra
authors.forename: Swagat Kumar
authors.initials: SK
authors.lastname: Patra
authors.forename: Rudra Prasanna
authors.initials: RP
authors.lastname: Panda
authors.forename: Ramya
authors.initials: R
authors.lastname: Mohanta
authors.forename: Ashis
authors.initials: A
authors.lastname: Saha
authors.forename: Jatindra Nath
authors.initials: JN
authors.lastname: Saha
authors.forename: Kanta
authors.initials: K
authors.lastname: Das Mahapatra
authors.forename: Pallipuram
authors.initials: P
authors.lastname: Jayasankar
authors.forename: Hirak Kumar
authors.initials: HK
authors.lastname: Barman
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D002347:Carps / Q000235:genetics* / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis* / Q000235:genetics; D011401:Promoter Regions, Genetic; D029961:Zebrafish Proteins
chemical_list: D050814:Octamer Transcription Factor-3; C085437:Pou5f3 protein, zebrafish; D029961:Zebrafish Proteins