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© The Rockefeller University Press,
0021-9525/1997//1851 $5.00
The Journal of Cell Biology, Volume 139, Number 7,
, 1997 1851-1859
Article |
A Novel Family of Serine/Threonine Kinases Participating in Spermiogenesis
Institute of Anatomy, CH-3012 Berne, Switzerland; Institute of Veterinary Parasitology, CH-3012 Berne, Switzerland; and || Department of Urology, Inselspital, CH-3010 Berne, Switzerland
The molecular mechanisms regulating the spectacular cytodifferentiation observed during spermiogenesis are poorly understood. We have recently identified a murine testis-specific serine kinase (tssk) 1, constituting a novel subfamily of serine/threonine kinases. Using low stringency screening we have isolated and molecularly characterized a second closely related family member, tssk 2, which is probably the orthologue of the human DGS-G gene. Expression of tssk 1 and tssk 2 was limited to the testis of sexually mature males. Immunohistochemical staining localized both kinases to the cytoplasm of late spermatids and to structures resembling residual bodies. tssk 1 and tssk 2 were absent in released sperms in the lumen of the seminiferous tubules and the epididymis, demonstrating a tight window of expression restricted to the last stages of spermatid maturation. In vitro kinase assays of immunoprecipitates containing either tssk 1 or tssk 2 revealed no autophosphorylation of the kinases, however, they led to serine phosphorylation of a coprecipitating protein of
65 kD. A search for interacting proteins using the yeast two-hybrid system with tssk 1 and tssk 2 cDNA as baits and a prey cDNA library from mouse testis, led to the isolation of a novel cDNA, interacting specifically with both tssk 1 and tssk 2, and encoding the coprecipitated 65-kD protein phosphorylated by both kinases. Interestingly, expression of the interacting clone was also testis specific and paralleled the developmental expression observed for the kinases themselves. These results represent the first demonstration of the involvement of a distinct kinase family, the tssk serine/threonine kinases, together with a substrate in the cytodifferentiation of late spermatids to sperms.
Abbreviations used in this paper: nt, nucleotides; pBS, pBluescript; poly(A), polyadenylated; SC, selection medium; SSC, standard saline citrate; tssk 1 and 2, testis-specific serine kinase 1 and 2.
Address all correspondence to A. Ziemiecki, Department of Clinical Research, Tiefenaustrasse 120, CH-3004 Berne, Switzerland. Tel: 41-31-308-8014. Fax: 41-31-308-8028. E-mail: andrew.ziemiecki{at}dkf3.unibe.ch
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