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Rabbit Epididymal Secretory Proteins. III. Molecular Cloning and Characterization of the Complementary DNA for REP38¹

^a Pest Animal Control Cooperative Research Centre, CSIRO Sustainable Ecosystems, Canberra, ACT 2601, Australia ^b Department of Biological Sciences, University of Newcastle, Callaghan, NSW 2308, Australia

REP38 is a rabbit epididymal secretory protein of 38 kDa that has recently been shown to interact with spermatozoa. A rabbit epididymal cDNA expression library was screened with a polyclonal antibody raised against REP38. A single clone (REP38-c1) with an open reading frame encoding a polypeptide of 666 amino acids was obtained. Cleavage of a 22-amino acid N-terminal signal peptide revealed a mature protein with a theoretical molecular mass of 74.5 kDa. Northern blot analysis revealed the presence of two cross-hybridizing transcripts of approximately 1.3 and 2.5 kilobases that appear to result from alternative mRNA splicing. This finding may explain the discrepancies between the observed (38 kDa) and deduced molecular mass of REP38. Expression of both transcripts was epididymis specific and was detected only in regions 2–6. During development, the expression of REP38-c1 mRNA was initiated between 1 and 2 mo postnatum and therefore precedes the appearance of sperm within the lumen of the epididymis. These findings are in agreement with the immunohistochemical localization of the REP38 protein. Androgen deprivation induced by orchidectomy reduced REP38-c1 mRNA levels below the limit of detection, an effect that was reversed by administration of exogenous testosterone. Although REP38-c1 mRNA was detected only in the rabbit epididymis, database searches indicated homology with two rat testis specific cDNAs, KTT4 and odf2, which encode sperm outer dense fiber proteins.

First decision: 24 May 1999.

¹ This work was supported by a grant from the Australian Research Council, the Research Management Committee, University of Newcastle, and the Pest Animal Control Cooperative Research Centre, Canberra, Australia. B.N. was supported by an Australian Postgraduate Award and a Pest Animal Control Cooperative Research Centre Student Award.

² Correspondence: Michael Holland, Pest Animal Control Cooperative Research Centre, CSIRO Sustainable Ecosystems, P.O. Box 284, Canberra, ACT 2601, Australia. FAX: 61 2 6242 1511;michael.holland{at}csiro.au

This article has been cited by other articles:

				B. Nixon, R. C. Jones, and M. K. Holland Molecular and Functional Characterization of the Rabbit Epididymal Secretory Protein 52, REP52 Biol Reprod, May 1, 2008; 78(5): 910 - 920. [Abstract] [Full Text] [PDF]