HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 69/1/254    most recent biolreprod.102.010975v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Thompson, W. E. Articles by Sutovsky, P. Search for Related Content PubMed PubMed Citation Articles by Thompson, W. E. Articles by Sutovsky, P. Agricola Articles by Thompson, W. E. Articles by Sutovsky, P.

Ubiquitination of Prohibitin in Mammalian Sperm Mitochondria: Possible Roles in the Regulation of Mitochondrial Inheritance and Sperm Quality Control¹

Departments of Obstetrics and Gynecology and Cooperative Reproductive Science Research Center, Morehouse School of Medicine,³ Atlanta, Georgia 30310 Department of Zoology, Center for Neuroscience and Cell Biology, University of Coimbra,⁴ Largo Marques de Pombal, 3004-517 Coimbra, Portugal Departments of Animal Sciences and Obstetrics and Gynecology, University of Missouri-Columbia,⁵ Columbia, Missouri 65211-5300

Ubiquitination of the sperm mitochondria during spermatogenesis has been implicated in the targeted degradation of paternal mitochondria after fertilization, a mechanism proposed to promote the predominantly maternal inheritance of mitochondrial DNA in humans and animals. The identity of ubiquitinated substrates in the sperm mitochondria is not known. In the present study, we show that prohibitin, a highly conserved, 30- to 32-kDa mitochondrial membrane protein, occurs in a number of unexpected isoforms, ranging from 64 to greater than 185 kDa in the mammalian sperm mitochondria, which are the ubiquitinated substrates. These bands bind antiubiquitin antibodies, displaying a pattern consistent with polyubiquitinated "ladders." Immunoprecipitation of sperm extracts with antiprohibitin antibodies followed by probing of the resultant immunocomplexes with antiubiquitin yields a banding pattern identical to that observed by antiprohibitin Western blot analysis. In fact, the presumably nonubiquitinated 30-kDa prohibitin band shows no antiubiquitin immunoreactivity. We demonstrate that ubiquitination of prohibitin occurs in testicular spermatids and spermatozoa. Ubiquitinated prohibitin molecules also accumulate in the defective fractions of ejaculated spermatozoa, which are thought to undergo surface ubiquitination during epididymal passage. In such sperm fractions, ubiquitin also coprecipitates with tubulin and microtubule-associated proteins, presumably contributed by the axonemes of defective, ubiquitinated spermatozoa. The results of the present study suggest that prohibitin is one of the ubiquitinated substrates that makes the sperm mitochondria recognizable by the egg's ubiquitin-proteasome dependent proteolytic machinery after fertilization and most likely facilitates the marking of defective spermatozoa in the epididymis for degradation.

¹ Supported by the Food for the 21st Century Program of the University of Missouri-Columbia and by U.S. Department of Agriculture (2002-02069; 99-35203-7785) and NIH/NIOSH (OH07324-01) grants to P.S. W.E.T. was supported by NIH (HD41749, GM08248, RR03034, and NCI P50-CA83591 SPORE in Ovarian Cancer). J.R.-S. was supported by a grant from FCT, Portugal (POCTI/ESP/38049/2001).

² Correspondence: Peter Sutovsky, Assistant Professor, University of Missouri-Columbia, S141 ASRC, 920 East Campus Drive, Columbia, MO 65211-5300. FAX: 573 884 5540; sutovskyp{at}missouri.edu

This article has been cited by other articles:

				J. N. Wolff and N. J. Gemmell Estimating Mitochondrial DNA Content of Chinook Salmon Spermatozoa Using Quantitative Real-Time Polymerase Chain Reaction Biol Reprod, August 1, 2008; 79(2): 247 - 252. [Abstract] [Full Text] [PDF]

				M. M.J. Cohen, G. P. Leboucher, N. Livnat-Levanon, M. H. Glickman, and A. M. Weissman Ubiquitin-Proteasome-dependent Degradation of a Mitofusin, a Critical Regulator of Mitochondrial Fusion Mol. Biol. Cell, June 1, 2008; 19(6): 2457 - 2464. [Abstract] [Full Text] [PDF]

				R. Maor, A. Jones, T. S. Nuhse, D. J. Studholme, S. C. Peck, and K. Shirasu Multidimensional Protein Identification Technology (MudPIT) Analysis of Ubiquitinated Proteins in Plants Mol. Cell. Proteomics, April 1, 2007; 6(4): 601 - 610. [Abstract] [Full Text] [PDF]

				I. Chowdhury, W. Xu, J. K. Stiles, A. Zeleznik, X. Yao, R. Matthews, K. Thomas, and W. E. Thompson Apoptosis of Rat Granulosa Cells after Staurosporine and Serum Withdrawal Is Suppressed by Adenovirus-Directed Overexpression of Prohibitin Endocrinology, January 1, 2007; 148(1): 206 - 217. [Abstract] [Full Text] [PDF]

				X. Peng, R. Mehta, S. Wang, S. Chellappan, and R. G. Mehta Prohibitin is a novel target gene of vitamin d involved in its antiproliferative action in breast cancer cells. Cancer Res., July 15, 2006; 66(14): 7361 - 7369. [Abstract] [Full Text] [PDF]

				D. C. Logan The mitochondrial compartment J. Exp. Bot., March 1, 2006; 57(6): 1225 - 1243. [Abstract] [Full Text] [PDF]

				K. Mitra, N. Rangaraj, and S. Shivaji Novelty of the Pyruvate Metabolic Enzyme Dihydrolipoamide Dehydrogenase in Spermatozoa: CORRELATION OF ITS LOCALIZATION, TYROSINE PHOSPHORYLATION, AND ACTIVITY DURING SPERM CAPACITATION J. Biol. Chem., July 8, 2005; 280(27): 25743 - 25753. [Abstract] [Full Text] [PDF]

				A. J. Whitworth, D. A. Theodore, J. C. Greene, H. Benes, P. D. Wes, and L. J. Pallanck Increased glutathione S-transferase activity rescues dopaminergic neuron loss in a Drosophila model of Parkinson's disease PNAS, May 31, 2005; 102(22): 8024 - 8029. [Abstract] [Full Text] [PDF]

				A. Sharma and A. Qadri Vi polysaccharide of Salmonella typhi targets the prohibitin family of molecules in intestinal epithelial cells and suppresses early inflammatory responses PNAS, December 14, 2004; 101(50): 17492 - 17497. [Abstract] [Full Text] [PDF]

				W. E. Thompson, E. Asselin, A. Branch, J. K. Stiles, P. Sutovsky, L. Lai, G.-S. Im, R. S. Prather, S. C. Isom, E. Rucker III, et al. Regulation of Prohibitin Expression During Follicular Development and Atresia in the Mammalian Ovary Biol Reprod, July 1, 2004; 71(1): 282 - 290. [Abstract] [Full Text] [PDF]