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BOR - Papers in Press, published online ahead of print March 21, 2007.
Biol Reprod 2007, 10.1095/biolreprod.106.057166
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Submitted September 5, 2006
Returned for revision October 27, 2006
Accepted March 20, 2007

Gamete Biology


Cysteine-Rich Secretory Protein 2 Binds to Mitogen-Activated Protein Kinase Kinase Kinase 11 in Mouse Sperm

Gerard M. Gibbs , Deborah M. Bianco , Duangporn Jamsai , Amy Herlihy , Sika Ristevski , R. John Aitken , David M. de Kretser , and Moira K. O'Bryan *

* To whom correspondence should be addressed. E-mail: moira.obryan{at}med.monash.edu.au.

Abstract
CRISP2 (previously TPX1) is a testis-enriched member of the cysteine-rich secretory protein (CRISP) family and has been localized to both the sperm acrosome and tail. Like all members of the mammalian CRISP family its expression pattern is strongly suggestive of a role in male fertility, but functional support for this hypothesis remains limited. In order to determine the biochemical pathways within which CRISP2 is a component, the putative mature form of CRISP2 was used as bait in a yeast two hybrid screen of a mouse testis expression library. One of the most frequently identified interacting partners was mitogen activated protein kinase kinase kinase 11 (MAP3K11). Sequencing and deletion experiments showed that the carboxyl-most 20 amino acids of MAP3K11 interacted with the CRISP domain of CRISP2. This interaction was confirmed using pull down experiments and the cellular context supported by the localization of CRISP2 and MAP3K11 to the acrosome of the developing spermatids and epididymal spermatozoa. Interestingly, mouse epididymal sperm contained a ~60 kDa variant of MAP3K11 which may be a result of proteolytic cleavage of the longer 93 kDa form seen in many tissues. These data raise the possibility that CRISP2 is a MAP3K11 modifying protein, or alternatively that MAP3K11 acts to phosphorylate CRISP2 during acrosome development.

Key words: Male Reproductive Tract • Testis • Sperm • Sperm maturation • Spermatogenesis


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D. Jamsai, A. Reilly, S.J. Smith, G.M. Gibbs, H.W.G. Baker, R.I. McLachlan, D.M. de Kretser, and M.K. O'Bryan
Polymorphisms in the human cysteine-rich secretory protein 2 (CRISP2) gene in Australian men
Hum. Reprod., September 1, 2008; 23(9): 2151 - 2159.
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D. Jamsai, D. M Bianco, S. J Smith, D. J Merriner, J. D Ly-Huynh, A. Herlihy, B. Niranjan, G. M Gibbs, and M. K O'Bryan
Characterization of gametogenetin 1 (GGN1) and its potential role in male fertility through the interaction with the ion channel regulator, cysteine-rich secretory protein 2 (CRISP2) in the sperm tail
Reproduction, June 1, 2008; 135(6): 751 - 759.
[Abstract] [Full Text] [PDF]


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