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BOR - Papers in Press, published online ahead of print September 14, 2005.
Biol Reprod 2005, 10.1095/biolreprod.105.045484
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BIOLOGY OF REPRODUCTION 74, 102–108 (2006)
DOI: 10.1095/biolreprod.105.045484
© 2006 by the Society for the Study of Reproduction, Inc.

Research Article

Cullin3 Is a KLHL10-Interacting Protein Preferentially Expressed During Late Spermiogenesis

Shouhua Wang, Huili Zheng, Yumiko Esaki, Fiona Kelly, and Wei Yan 1

Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, Nevada 89557

ABSTRACT

Kelch-like 10 (KLHL10) is a member of the BTB (Bric-a-brac, Tramtrack, and Broad-Complex)-kelch protein superfamily essential for spermiogenesis and male fertility. In a search for KLHL10-interacting proteins using a yeast two-hybrid assay, we identified Cullin3 (CUL3) as one of multiple KLHL10-interacting partners. Yeast cotransformation assays revealed that CUL3 bound the BTB/POZ domain of KLHL10. Northern blot and quantitative RT-PCR analyses demonstrated that Cul3 mRNA was preferentially expressed in the testis. In situ hybridization analysis localized Cul3 mRNA to spermatids in the adult testis. CUL3 protein was detected in elongating and elongated spermatids (steps 10–16) by immunofluorescent microscopy. The expression pattern of CUL3 resembles KLHL10. CUL3 was coimmunoprecipated with KLHL10, and KLHL10 was also detected in the CUL3 immunoprecipitants using testis lysates. These findings suggest that KLHL10, like other BTB/kelch proteins, interacts with CUL3 to form a CUL3-based ubiquitin E3 ligase that functions specifically in the testis to mediate protein ubiquitination during spermiogenesis.

BTB protein, Cullin3, infertility, spermatid, spermatogenesis, testis, Ubiquitin E3 ligase, ubiquitination

FOOTNOTES

1 Correspondence: Wei Yan, Department of Physiology and Cell Biology, University of Nevada School of Medicine, MS352, 1664 North Virginia Street, Reno, NV 89557. FAX: 775 784 6903; weiyan{at}unr.edu






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Copyright © 2006 by the Society for the Study of Reproduction.