Journal of Cell Science, Vol 113, Issue 1 103-112, Copyright © 2000 by Company of Biologists

JOURNAL ARTICLES

Ropporin, a sperm-specific binding protein of rhophilin, that is localized in the fibrous sheath of sperm flagella

A Fujita, K Nakamura, T Kato, N Watanabe, T Ishizaki, K Kimura, A Mizoguchi and S Narumiya
Department of Pharmacology and Department of Anatomy and Neurobiology, Kyoto University Faculty of Medicine, Sakyo, Kyoto 606-8501, Japan.

The small GTPase Rho; functions as a molecular switch that regulates various cellular processes such as cell adhesion, motility, gene expression and cytokinesis. We previously isolated several putative Rho; targets including rhophilin which bound selectively to the GTP-bound form of Rho;. Rhophilin is expressed highly in testis and is localized specifically in sperm flagella. The presence of a PDZ domain at the carboxy terminus of rhophilin suggested that rhophilin works as an adaptor molecule. To test this hypothesis, we employed a yeast two hybrid system using the rhophilin PDZ domain as a bait, and screened a mouse testis cDNA library. We isolated several positive clones containing the same insert. The open reading frame of the cDNA encoded a novel protein of 212 amino acids designated as ropporin from a Japanese word 'oppo' (the tail). The amino-terminal 40 amino acid sequence of ropporin showed high homology to that of the regulatory subunit of type II cAMP-dependent protein kinase, which is involved in dimerization and binding to A-kinase anchoring proteins. Consistently, a yeast two hybrid assay and gel filtration of recombinant ropporin indicated that ropporin dimerizes through this domain. Deletion analysis indicated that the carboxy-terminal four amino acids are essential for binding of ropporin to rhophilin, and ropporin and RhoV14 coprecipitated in the presence of rhophilin in vitro. Northern blot analysis showed that ropporin is exclusively expressed in testis, and induced at the late stage of spermatogenesis. This induction paralleled that of rhophilin. Immunocytochemistry using anti-ropporin antibody showed that ropporin is localized in the principal piece and the end piece of sperm flagella. Electronmicroscopy revealed that ropporin is mostly localized in the inner surface of the fibrous sheath while rhophilin is present in the outer surface of the outer dense fiber. These results suggest that rhophilin and ropporin may form a complex in sperm flagella.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				S. E Fiedler, M. Bajpai, and D. W Carr Identification and Characterization of RHOA-Interacting Proteins in Bovine Spermatozoa Biol Reprod, January 1, 2008; 78(1): 184 - 192. [Abstract] [Full Text] [PDF]

				L. M Correa, A. Thomas, and S. A Meyers The Macaque Sperm Actin Cytoskeleton Reorganizes in Response to Osmotic Stress and Contributes to Morphological Defects and Decreased Motility Biol Reprod, December 1, 2007; 77(6): 942 - 953. [Abstract] [Full Text] [PDF]

				Y.-F. Li, W. He, K. N. Jha, K. Klotz, Y.-H. Kim, A. Mandal, S. Pulido, L. Digilio, C. J. Flickinger, and J. C. Herr FSCB, a Novel Protein Kinase A-phosphorylated Calcium-binding Protein, Is a CABYR-binding Partner Involved in Late Steps of Fibrous Sheath Biogenesis J. Biol. Chem., November 23, 2007; 282(47): 34104 - 34119. [Abstract] [Full Text] [PDF]

				D. Escalier Knockout mouse models of sperm flagellum anomalies Hum. Reprod. Update, July 1, 2006; 12(4): 449 - 461. [Abstract] [Full Text] [PDF]

				C. Yang and P. Yang The Flagellar Motility of Chlamydomonas pf25 Mutant Lacking an AKAP-binding Protein Is Overtly Sensitive to Medium Conditions Mol. Biol. Cell, January 1, 2006; 17(1): 227 - 238. [Abstract] [Full Text] [PDF]

				L. Blumenstein and M. R. Ahmadian Models of the Cooperative Mechanism for Rho Effector Recognition: IMPLICATIONS FOR RhoA-MEDIATED EFFECTOR ACTIVATION J. Biol. Chem., December 17, 2004; 279(51): 53419 - 53426. [Abstract] [Full Text] [PDF]

				M. H. Modarressi, M. Cheng, H. A. Tarnasky, N. Lamarche-Vane, D. G. de Rooij, Y. Ruan, and F. A. van der Hoorn A Novel Testicular RhoGAP-Domain Protein Induces Apoptosis Biol Reprod, December 1, 2004; 71(6): 1980 - 1990. [Abstract] [Full Text] [PDF]

				M. H. Modarressi, B. Behnam, M. Cheng, K. E. Taylor, J. Wolfe, and F. A. van der Hoorn Tsga10 Encodes a 65-Kilodalton Protein That Is Processed to the 27-Kilodalton Fibrous Sheath Protein Biol Reprod, March 1, 2004; 70(3): 608 - 615. [Abstract] [Full Text] [PDF]

				A. Miranda-Vizuete, K. Tsang, Y. Yu, A. Jimenez, M. Pelto-Huikko, C. J. Flickinger, P. Sutovsky, and R. Oko Cloning and Developmental Analysis of Murid Spermatid-specific Thioredoxin-2 (SPTRX-2), a Novel Sperm Fibrous Sheath Protein and Autoantigen J. Biol. Chem., November 7, 2003; 278(45): 44874 - 44885. [Abstract] [Full Text] [PDF]

				R. M. Turner Tales From the Tail: What Do We Really Know About Sperm Motility? J Androl, November 1, 2003; 24(6): 790 - 803. [Full Text] [PDF]

				S. Hayashi, J. Yang, L. Christenson, R. Yanagimachi, and N. B. Hecht Mouse Preimplantation Embryos Developed from Oocytes Injected with Round Spermatids or Spermatozoa Have Similar but Distinct Patterns of Early Messenger RNA Expression Biol Reprod, October 1, 2003; 69(4): 1170 - 1176. [Abstract] [Full Text] [PDF]

				P. R. Brown, K. Miki, D. B. Harper, and E. M. Eddy A-Kinase Anchoring Protein 4 Binding Proteins in the Fibrous Sheath of the Sperm Flagellum Biol Reprod, June 1, 2003; 68(6): 2241 - 2248. [Abstract] [Full Text] [PDF]

				J. W. Peck, M. Oberst, K. B. Bouker, E. Bowden, and P. D. Burbelo The RhoA-binding protein, Rhophilin-2, Regulates Actin Cytoskeleton Organization J. Biol. Chem., November 8, 2002; 277(46): 43924 - 43932. [Abstract] [Full Text] [PDF]

				Y. Yu, R. Oko, and A. Miranda-Vizuete Developmental Expression of Spermatid-Specific Thioredoxin-1 Protein: Transient Association to the Longitudinal Columns of the Fibrous Sheath During Sperm Tail Formation Biol Reprod, November 1, 2002; 67(5): 1546 - 1554. [Abstract] [Full Text] [PDF]

				M. D. Anway, N. Ravindranath, M. Dym, and M. D. Griswold Identification of a Murine Testis Complementary DNA Encoding a Homolog to Human A-Kinase Anchoring Protein-Associated Sperm Protein Biol Reprod, June 1, 2002; 66(6): 1755 - 1761. [Abstract] [Full Text] [PDF]

				M. Eda, S. Yonemura, T. Kato, N. Watanabe, T. Ishizaki, P. Madaule, and S. Narumiya Rho-dependent transfer of Citron-kinase to the cleavage furrow of dividing cells J. Cell Sci., March 11, 2002; 114(18): 3273 - 3284. [Abstract] [Full Text] [PDF]

				E. de Lamirande and C. Gagnon The extracellular signal-regulated kinase (ERK) pathway is involved in human sperm function and modulated by the superoxide anion Mol. Hum. Reprod., February 1, 2002; 8(2): 124 - 135. [Abstract] [Full Text] [PDF]

				H. M. Lacy and R. D. Sanderson Sperm protein 17 is expressed on normal and malignant lymphocytes and promotes heparan sulfate-mediated cell-cell adhesion Blood, October 1, 2001; 98(7): 2160 - 2165. [Abstract] [Full Text] [PDF]

				R. D. Catalano, E. W. Hillhouse, and M. Vlad Developmental Expression and Characterization of FS39, a Testis Complementary DNA Encoding an Intermediate Filament-Related Protein of the Sperm Fibrous Sheath Biol Reprod, July 1, 2001; 65(1): 277 - 287. [Abstract] [Full Text] [PDF]

				Y. Nakagawa, Y. Yamane, T. Okanoue, S. Tsukita, and S. Tsukita Outer Dense Fiber 2 Is a Widespread Centrosome Scaffold Component Preferentially Associated with Mother Centrioles: Its Identification from Isolated Centrosomes Mol. Biol. Cell, June 1, 2001; 12(6): 1687 - 1697. [Abstract] [Full Text] [PDF]

				T Kato, N Watanabe, Y Morishima, A Fujita, T Ishizaki, and S Narumiya Localization of a mammalian homolog of diaphanous, mDia1, to the mitotic spindle in HeLa cells J. Cell Sci., January 2, 2001; 114(4): 775 - 784. [Abstract] [PDF]

				D. W. Carr, A. Fujita, C. L. Stentz, G. A. Liberty, G. E. Olson, and S. Narumiya Identification of Sperm-specific Proteins That Interact with A-kinase Anchoring Proteins in a Manner Similar to the Type II Regulatory Subunit of PKA J. Biol. Chem., May 11, 2001; 276(20): 17332 - 17338. [Abstract] [Full Text] [PDF]