Testis-Specific RT7 Protein Localizes to the Sperm Tail and Associates with Itself1

Higgy, N A; Pastoor, Tina E.; Renz, Christian; Tarnasky, Heide A.; Hoorn, Frans A. van der

doi:10.1095/biolreprod50.6.1357

Cited by 26 publications

(20 citation statements)

References 21 publications

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“…Izumo sperm-egg fusion 1 (Izumo1) (11) and AKAP82, the major protein of the fibrous sheath of the sperm flagellum (12), were present in the membrane and FS fractions, respectively. Odf1, the major ODF protein (13), was extracted only with urea (Fig. 5B).…”

Section: Mca Homozygous Mutant Malesmentioning

confidence: 99%

Meichroacidin Containing the Membrane Occupation and Recognition Nexus Motif Is Essential for Spermatozoa Morphogenesis

Tokuhiro

Hirose

Miyagawa

et al. 2008

Journal of Biological Chemistry

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Meichroacidin (MCA) is a highly hydrophilic protein that contains the membrane occupation and recognition nexus motif. MCA is expressed during the stages of spermatogenesis from pachytene spermatocytes to mature sperm development and is localized in the male meiotic metaphase chromosome and sperm flagellum. MCA sequences are highly conserved in Ciona intestinalis, Cyprinus carpio, and mammals. To investigate the physiological role of MCA, we generated MCA-disrupted mutant mice; homozygous MCA mutant males were infertile, but females were not. Sperm was rarely observed in the caput epididymidis of MCA mutant males. However, little to no difference was seen in testis mass between wild-type and mutant mice. During sperm morphogenesis, elongated spermatids had retarded flagellum formation and might increase phagocytosis by Sertoli cells. Immunohistochemical analysis revealed that MCA interacts with proteins located on the outer dense fibers of the flagellum. The testicular sperm of MCA mutant mice was capable of fertilizing eggs successfully via intracytoplasmic sperm injection and generated healthy progeny. Our results suggest that MCA is essential for sperm flagellum formation and the production of functional sperm.

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Section: Mca Homozygous Mutant Malesmentioning

confidence: 99%

Meichroacidin Containing the Membrane Occupation and Recognition Nexus Motif Is Essential for Spermatozoa Morphogenesis

Tokuhiro

Hirose

Miyagawa

et al. 2008

Journal of Biological Chemistry

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“…Immunoelectron microscopy with the mAb showed that, in mature sperm, KLC3 associates with the surface of ODFs and is present in the space between adjacent ODFs and between mitochondria and ODFs, which are devoid of tubulins (38). KLC3 is not present in the medulla of ODFs, indicating that KLC3 is not an integral ODF protein like ODF1 (36,54) and ODF2 (29,55). Indeed, Western blot analysis of highly purified ODFs failed to detect KLC3 (Fig.…”

Section: Table I Klc3 Binding To Odf1 In Yeastmentioning

confidence: 96%

“…Proteins from fractionated male germ cells obtained by centrifugal elutriation (36,37) or from ODFs and the FS were separated on 10 -15% gradient SDS-polyacrylamide gels and transferred to NitroPlus membranes. Membranes were blocked and washed as described above and were then incubated in Western blot binding buffer (30) containing 1 mM ATP and 1000 cpm/ml 35 S-labeled KLC3 protein, produced by in vitro translation, for 16 h at 4°C.…”

Section: Klc3-odf Binding Assaysmentioning

confidence: 99%

Association of Kinesin Light Chain with Outer Dense Fibers in a Microtubule-independent Fashion

Bhullar

Zhang

Junco

et al. 2003

Journal of Biological Chemistry

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Conventional kinesin I motor molecules are heterotetramers consisting of two kinesin light chains (KLCs) and two kinesin heavy chains. The interaction between the heavy and light chains is mediated by the KLC heptad repeat (HR), a leucine zipper-like motif. Kinesins bind to microtubules and are involved in various cellular functions, including transport and cell division. We recently isolated a novel KLC gene, klc3. klc3 is the only known KLC expressed in post-meiotic male germ cells. A monoclonal anti-KLC3 antibody was developed that, in immunoelectron microscopy, detects KLC3 protein associated with outer dense fibers (ODFs), unique structural components of sperm tails. No significant binding of KLC3 with microtubules was observed with this monoclonal antibody. In vitro experiments showed that KLC3-ODF binding occurred in the absence of kinesin heavy chains or microtubules and required the KLC3 HR. ODF1, a major ODF protein, was identified as the KLC3 binding partner. The ODF1 leucine zipper and the KLC3 HR mediated the interaction. These results identify and characterize a novel interaction between a KLC and a non-microtubule macromolecular structure and suggest that KLC3 could play a microtubule-independent role during formation of sperm tails.

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“…The antibodies were affinity-purified by binding them to filterimmobilized MBP-18-8-1 fusion protein as described [14]. Affinity-purified anti-OIP1 antibodies were used 1) in Western blotting experiments to detect testicular OIP1 protein as described [12] using protein extracts isolated from testis, spermatozoa, heart, and smooth muscle; and 2) in indirect immunofluorescence analysis of formaldehyde-fixed, Triton-permeabilized rat epididymal spermatozoa using Cy-3 conjugated goat anti-rabbit sera as described [11,16].…”

Section: Oip1 Protein Analysismentioning

confidence: 99%

“…The second ODF1 structural feature is a C-terminal CysGly-Pro (CGP) repeat [16], also present in the Drosophila Mst(3) CGP gene family [17][18][19]. The Mst(3) gene family is composed of seven genes, five small genes of similar size and exon-intron structure (Mst87F, Mst84Da, Mst84Db, Mst84Dc, and Mst84Dd) and two tandemly ar-544 ZARSKY ET AL.…”

Section: Introductionmentioning

confidence: 99%

Novel RING Finger Protein OIP1 Binds to Conserved Amino Acid Repeats in Sperm Tail Protein ODF11

Zarsky¹,

Cheng²,

Hoorn³

2003

Biology of Reproduction

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Outer dense fibers (ODFs) and the fibrous sheath (FS) are unique structures of the mammalian sperm tail. Recently, progress has been made in the molecular cloning of ODF and FS proteins, and because of this, questions addressing the morphogenesis and underlying protein network that make up sperm tail structures and their function can now be addressed. Using the N-terminal leucine zipper motif of the major ODF protein ODF1, we had previously isolated interacting proteins Odf2, Spag4, and Spag5. We report here a yeast two-hybrid strategy to isolate a novel rat testicular protein, OIP1, that binds to the evolutionarily conserved Cys-Gly-Pro repeats in the C-terminus of ODF1. OIP1 is expressed in round spermatids as well as in spermatocytes and several somatic tissues, albeit at a lower level. No expression was detectable in epididymis, heart, and smooth muscle. OIP1 protein localizes to the sperm tail in a pattern expected for an ODF1-interacting protein. OIP1 belongs to the family of RING finger proteins of the H2 subclass. Deletion of the putative RING motif significantly decreased binding to ODF1. Genomic analysis of rat Oip1 and Oip1 homologs indicates that Oip1 is highly conserved. Oip1 is subject to differential splicing and alternative polyadenylation events. It is interesting that Oip1 mRNAs have been reported that lack the exon encoding the putative RING finger.

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Testis-Specific RT7 Protein Localizes to the Sperm Tail and Associates with Itself1

Cited by 26 publications

References 21 publications

Meichroacidin Containing the Membrane Occupation and Recognition Nexus Motif Is Essential for Spermatozoa Morphogenesis

Meichroacidin Containing the Membrane Occupation and Recognition Nexus Motif Is Essential for Spermatozoa Morphogenesis

Association of Kinesin Light Chain with Outer Dense Fibers in a Microtubule-independent Fashion

Novel RING Finger Protein OIP1 Binds to Conserved Amino Acid Repeats in Sperm Tail Protein ODF11

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