Molecular Human Reproduction, Vol. 8, No. 6, 525-530,
June 2002
© 2002 European Society of Human Reproduction and Embryology
Testis and spermatogenesis |
Cloning and characterization of the human tektin-t gene
1 Department of Science for Laboratory Animal Experimentation, Research Institute for Microbial Diseases, Osaka University, Yamadaoka, 2 Department of Urology, Osaka University Medical School, Suita City, Osaka 565-0871 and 3 Otsuka GEN Research Institute, Otsuka Pharmaceutical Co., Ltd, Kagasuno Kawauchi-cho, Tokushima 771-0192, Japan
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Abstract |
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Tektin-t is a part of the tektin protein family, the members of which form filamentous polymers in the walls of ciliary and flagellar microtubules. In mice, tektin-t protein has been localized to the tail of mature sperm, suggesting that it has a role in the formation of sperm flagella and/or sperm motility. In the present study, we have cloned a human orthologue of mouse haploid germ cell-specific tektin-t. The cloned human cDNA and the predicted amino acid sequence showed 82 and 83% identity with mouse tektin-t respectively. Included were a sequence conserved in the tektin B1 family, the TEKTIN2 motif, and the consensus sequence in the tektin protein family composed of nona-peptide. Northern blot analysis of mRNA isolated from various human organs showed that the transcript was
1.7 kb in length and strongly expressed in the testis. Human (h-)tektin-t protein, having a molecular weight of 54 kDa, was exclusively expressed in the testis, whereas two additional stronger bands of 46 and 56 kDa existed in sperm. The h-tektin-t localized specifically to the principal piece of flagella and to the post-acrosomal region. The h-tektin-t gene was assigned to chromosome 1 by a radiation hybrid mapping technique. cDNA/gene mapping/microtubule/sperm/tektin
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Introduction |
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TopAbstractIntroductionMaterials and methodsResultsDiscussionAcknowledgementsReferences |
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In animal species, male gametes differentiate into sperm with extensive morphological and physiological changes. In mammals, haploid germ cell differentiation occurs continuously in the testis after puberty and sperm are produced throughout adult life (Russell et al., 1990
). Mature sperm are required to travel a long way for fertilization. The flagellum acts as the specialized apparatus to provide the locomotive force to enable sperm to reach the oocyte and also for actual penetration of the oocyte. One explanation for low fertility is that few sperm reach the oocyte due to poor migratory ability (Harpar, 1994; Froman et al., 1997; Donoghue et al., 1999). In fact, impairment of sperm movement is known to be a cause of male infertility (Jaffe and Oates, 1997). Thus, the understanding of the principal filamentous structures of the sperm tail and their physiological function in fertilization is important.
Tektins are the constitutive proteins of microtubules in cilia, flagella, basal bodies and centrioles (Linck et al., 1985; Steffen and Linck, 1988; Norrander et al., 1998; Larsson et al., 2000) and were originally isolated from sea urchins as a set of proteins, tektin A, B and C (57, 52 and 47 kDa respectively) (Linck et al., 1982; Norrander et al., 1996). Tektin A and B are thought to form core protofilaments of tektin filaments, and tektin C is thought to form homodimers assembled onto the periphery of these core protofilaments or to form a second separate tektin filament (Pirner and Linck, 1994). Tektins may function to provide stability and structural complexity to axonemal microtubules and work as templates and rulers in generating the three-dimensional organization of the axoneme (Linck and Langevin, 1982; Amos et al., 1986; Linck and Stephens, 1987; Norrander et al., 2000). Thus, tektins are thought to play a fundamental role in ciliary movement (Pirner et al., 1994; Nojima et al., 1995; Norrander et al., 1996).
We have previously isolated a new member of the tektin family, tektin-t, from a mouse haploid germ cell-specific cDNA library (Iguchi et al., 1999). Tektin-t is expressed exclusively in the testis, and is localized in flagella of mature sperm in mice. The predicted amino acid sequence of mouse tektin-t has the consensus sequence of tektin family proteins. Evolutionary conserved and constitutive proteins of sperm tail like tektin would play important roles in the formation and function of sperm flagella. Defects in the functions of such proteins may be associated with male infertility caused by an impairment of sperm movement such as asthenospermia or immotile sperm.
Here, we report the molecular cloning and chromosomal mapping of the human (h-) tektin-t gene encoding a protein localized to sperm flagella. During the review of this paper, similar results obtained by Wolkowicz et al. were published. These authors cloned a similar cDNA from a human testis cDNA library using degenerate oligonucleotides based on the DNA sequence of human sperm flagellar protein and named it h-tekt-B1 (Wolkowicz et al., 2002).
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Materials and methods |
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Cloning of human tektin-t cDNA
To isolate h-tektin-t, a human cDNA library constructed with plasmid vector pAP3neo was used (Tanaka et al., 1997
). More than 2x106 Escherichia coli containing recombinant plasmids were screened by hybridization with a 
-32P dCTP-labelled full-length cDNA of mouse tektin-t, prepared with a BcaBEST random primer kit (Takara, Shiga, Japan), for 20 h at 60°C, followed by two washes in a solution containing 2xsaline sodium citrate and 0.1% sodium dodecyl sulphate (SDS) at 55°C for 30 min each. Two independent positive clones were isolated, and the cDNA inserts were subcloned into pBluescript II vector, then sequenced by the dideoxy chain termination sequencing method (Sanger et al., 1997) with a Model 4000 sequencer (Li-COR, Lincoln, NE, USA) and analysed using the GenBank, EMBL, DDBJ, Swiss-Prot and PIR databases.
Northern blotting
Human multiple tissue Northern blots (Human MTN Blot II) were purchased from Clontech (Palo Alto, CA, USA). The filters were hybridized with a -32P dCTP-labelled cDNA according to the manufacturer's protocol.
Preparation of antiserum to h-tektin-t protein
The entire coding region of the h-tektin-t cDNA was PCR amplified with a primer set, forward (5'-GAAGATCTGGTTTCTGTGCCATGGCCA-3') and reverse (5'-CGGGATCCAAGGCTAGGCCAGCTCCAGCT-3'). The PCR products were digested by restriction enzymes (NcoI and BamHI) and subcloned into the pET30a expression vector (Novagen, Madison, WI, USA). The hexa–histidine-tagged recombinant protein was expressed in E.coli BL21 by induction with isopropyl-ß-D-thiogalactopyranoside and purified with His Bind Resin (Novagen) according to the manufacturer's instructions. Polyclonal antiserum was raised by immunization of Japanese white rabbits with histidine-tagged recombinant protein mixed with Freund adjuvant (Difco Laboratories, Detroit, MI, USA).
Western blotting
Freshly ejaculated human sperm obtained by masturbation were homogenized with Tris-buffered saline (TBS)–Tween 20 (TBS, 25 mmol/l Tris–HCl (pH 7.5), 150 mmol/l NaCl, and 50 mmol/l KCl, 0.05% Tween 20) on ice. After centrifugation, the supernatant was collected and the Bradford Protein Assay (Nacalai tesque, Kyoto, Japan) was conducted to estimate protein concentrations. Other protein samples (liver, lung, ovary, smooth muscle and testis) were purchased from Clontech. Aliquots of the samples (50 µg/lane) were then separated by 10% SDS–PAGE under reducing conditions (Laemmli, 1970), transferred to polyvinylidenedifluoride membrane filters (Millipore, Bedford, MA, USA) and incubated in Blocking Solution (Nacalai tesque). The filters were incubated with anti-h-tektin-t antiserum diluted 1000-fold with TBS. A horseradish peroxidase-conjugated anti-rabbit IgG antibody (Amersham Pharmacia Biotech, Tokyo, Japan) was used as a secondary antibody. Finally, the signals were detected by developing the filters with a POD staining kit (Wako, Osaka, Japan).
Lectin column chromatography of h-tektin-t
Freshly ejaculated human sperm were homogenized with a binding buffer [20 mmol/l Tris–HCl (pH 7.4), 0.5 mol/l NaCl, 1 mmol/l MnCl2 1 mmol/l MgCl2, and 1 mmol/l CaCl2]. After centrifugation, the supernatant was filtrated through a 0.45 µm pored SteraDisk (KURABO, Osaka, Japan). Concanavalin A sepharose (Amersham) was packed in a Poly-Prep Chromatography Column (BioRad, Hercules, CA, USA). After equilibration, the sample was loaded on the column. Bound substances were eluted with a linear gradient of methyl--D-glucoside (0, 0.1, 0.3 and 0.5 mol/l). Each fraction was collected in a plastic tube, precipitated with trichloroacetic acid, separated by 10% SDS–PAGE and analysed by Western blotting.
Immunofluorescence microscopy of human sperm
Human sperm samples from fertile male volunteers were separated from semen by washing with phosphate-buffered saline, and spotted onto Micro Slide Glass (Matsunami Glass, Osaka, Japan). The sperm samples were fixed with 100% EtOH for 10 min on ice. After blocking, the slides were incubated with anti-h-tektin-t rabbit antiserum (diluted 1:1000) overnight at 4°C and then treated with 5% normal donkey serum for 30 min at room temperature. After that the slides were incubated with fluorescein isothiocyanate-conjugated anti-rabbit IgG antibody (1:500; Amersham) for 2 h at room temperature. The samples were examined under a fluorescence microscope.
Radiation hybrid mapping
Radiation hybrid mapping (Watanabe et al., 1996) was carried out using the GeneBridge 4 Radiation Hybrid Panel (Research Genetics, Huntsville, AL, USA) according to the manufacturer's procedure, with a primer set (5'-AAGCTGACCGTGCCTGCTGAGAGGTTC-3' and 5'-ATTGCCCACCC-AACCCTGCCCTCCTC-3') designed from the 3' untranslated region of h-tektin-t cDNA. The amplification profile consisted of 30 cycles at 95°C for 30 s, 60°C for 30 s, and 72°C for 30 s.
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Results |
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Characterization of a cDNA encoding h-tektin-t
Sequence analysis of the cDNA clones revealed only an uninterrupted long open reading frame (nucleotides 1–1290), encoding 430 amino acid residues (GenBank accession number: AB033823) (Figure 1
). The h-tektin-t had a nearly identical sequence to that of human tektin-B1-like protein [GenBank/EMBL/DDBJ accession number: AF054910 (Wolkowicz et al., 2002)] with four nucleotide changes (969 C to T, 1044 C to T, 1046 C to T and 1442 G to A) and one amino acid change (329 L to F).
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A computer-assisted database search indicated that the predicted protein sequence possessed the TEKTIN 2 motif, RTYRPNVELCRDQAQYGLTDE, at amino acids 321–341. This motif contains the nona-peptide consensus sequence RPNVELCRD, conserved from sea urchin to human, and might have a crucial role in the function of tektins. A polyadenylation signal of h-tektin-t is located at nucleotides 1363–1368, similar to the position in mouse tektin-t (GenBank accession number: AB027138). The deduced amino acid sequence of h-tektin-t has five N-glycosylation sites (A.A. 47–50, 51–54, 60–63, 198–201 and 416–419). The h-tektin-t cDNA and its predicted amino acid sequence shows high homology with mouse tektin-t, 82 and 83% identity respectively.
Expression of h-tektin-t mRNA and protein
Approximately 1.7 kb long transcripts of h-tektin-t mRNA were expressed strongly in testis and weakly in ovary on human multiple tissue Northern blots (Figure 2). Other tissues showed no expression of h-tektin-t mRNA.
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Western blot analysis showed a single band of 54 kDa exclusively detected in human testis. In sperm, three bands were detectable; a major strong band of 56 kDa, a minor band of 46 kDa and a very faint band of 54 kDa (Figure 3
). Although a different molecular sized band was also detected in liver, it would be a cross-reactive material since no positive signal was observed in Northern blots of the liver.
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Lectin column chromatography of h-tektin-t
To study the cause of the difference in molecular weight between the forms in sperm and testis, glycosylation of h-tektin-t was examined, since five predicted N-glycosylation sites were present in the deduced amino acid sequence of h-tektin-t (Figure 1
). Concanavalin A (Con A) sepharose column chromatography of human sperm lysate showed that the larger h-tektin-t molecules (56 and 54 kDa) were specifically bound to the Con A column and eluted with a buffer containing hapten sugar (Figure 4), indicating that h-tektin-t of 56 and 54 kDa in sperm were N-glycosylated. In contrast h-tektin-t with a molecular weight of 46 kDa could not bind to the Con A column.
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Immunofluorescence microscopy of human sperm
Immunostaining for h-tektin-t was positive in the flagellum as in the case for mouse sperm, but the staining was not homogeneous; rather, it was discontinuous and punctuated. In contrast with mouse sperm, human sperm were stained in the post-acrosomal region of the head showing two positive spots in the tangential view (Figure 5
).
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Chromosome mapping of the h-tektin-t gene
The h-tektin-t gene was mapped to the short arm of chromosome 1, 0.1 cR telomeric of AFM094tb7 (LOD = 16.655) and 2.33 cR centromeric of AFMa107wf9 (LOD = 12.065) by using a GeneBridge 4 Radiation Hybrid panel (data not shown) and by two-point maximum-likelihood analysis software through the URL at http://www.genome.wi.mit.edu/cgi-bin/contig/rhmapper.pl. These markers lie in the region 1p34.3. Furthermore, a BLAST search in the human genome database of NCBI revealed that the h-tektin-t gene localized to chromosome 1pter-p32.3 (GenBank accession number: NT 004568.4), consistent with our data.
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Discussion |
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TopAbstractIntroductionMaterials and methodsResultsDiscussionAcknowledgementsReferences |
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Tektin family proteins have been studied for a long time, and the studies have shown that the tektins should contribute to the formation and reinforcement of ciliary and flagellar microtubules (Larsson et al., 2000
). The tektin-t was first isolated from a haploid germ cell-specific mouse cDNA library (Iguchi et al., 1999). Here, we report the molecular cloning of a human orthologue of tektin-t (h-tektin-t), which showed high homology with mouse tektin-t in both cDNA (82%) and deduced amino acid (83%) sequences. Amino acid sequences of both human and mouse tektin-t proteins contained a TEKTIN 2 motif, conserved also in sea urchin tektin B1 (Chen et al., 1993). The conservation of the TEKTIN 2 motif suggested that it has an important role in these molecules.
The h-tektin-t gene was mapped to chromosome 1 by radiation hybrid mapping, and a database search showed it to be a single gene located at 1pter-p32.3 with 10 exons (GenBank accession number: NT 004568.4) spread over 4.2 kbp. Recently, Olesen et al. mapped two transcripts differentially expressed in testis to 1q22-q23 and 1p36, and another transcript which was expressed ubiquitously but predominantly in testis to 1p32-p33 (Olesen et al., 2001). In addition, they indicated that three of five emphasized candidate genes for male infertility and 76 of 265 translocation breakpoints reported in infertile men are located on chromosome 1. These results suggested the presence on chromosome 1 of genes with a role in male fertility, and that the h-tektin-t gene might be a candidate for one of these genes.
The tektin-t transcripts of mouse and human were shown to be almost the same size, 1.7 kb, by Northern blot analysis. This analysis showed that h-tektin-t mRNA is exclusively expressed in testis and ovary; however, some transcripts were detected in embryonic organs by dot blot analysis of RNA (Wolkowicz et al., 2002). The predicted amino acid sequence and calculated molecular weights were also conserved in mouse and human tektin-t (49 kDa). Detailed analysis of the h-tektin-t protein gave us some different and additional results to the previous observation (Wolkowicz et al., 2002).
In Western blot analysis of human sperm, h-tektin-t was detected as major bands of 56 and 46 kDa, while in testis a single band of 54 kDa was detected, and this also existed as a faint band in the sperm fraction (Figure 3). Mammalian sperm undergo both protein and saccharide modifications, and a change in their phosphorylation profile during transit of the epididymis (Tulsiani et al., 1998; Nath and Majumder, 1999). The deduced amino acid sequence of h-tektin-t has five N-glycosylation sites. The presence of glycosyltransferases and glycosidases has been reported in rat and boar sperm (Tulsiani et al., 1998; Kuno et al., 2000). Also, a glycosidase, ß-D-glucuronidase, is reported to be associated with the sperm cytoskeleton (Lopes et al., 2001). We demonstrated here that the larger of the forms (56 and 54 kDa) of h-tektin-t in sperm bound to the Con A column chromatography, whereas the smaller h-tektin-t (46 kDa) did not. This indicated that the larger h-tektin-t (56 and 54 kDa) should be N-glycosylated (Gupta et al., 1996). Thus, h-tektin-t appears to be first glycosylated in the testis, with additional glycosylation as well as deglycosylation events occurring during epididymal maturation.
The N-linked oligosaccharide consists of a core structure, trimannoside and bi-N-acetylglucosamine, and side chain(s). Previous reports have described that Con A has enhanced affinity for a large N-linked oligomannose carbohydrate (Man5-9 glycopeptide) (Ogata et al., 1975; Kobata and Yamashita, 1993; Gupta et al., 1996). As the calculated molecular weight of one N-linked oligosaccharide (consisting of a core structure with a side chain of Man5-9) is 2.1– 2.8 kDa, one could predict that the h-tektin-t of 46 kDa is the non-glycosylated form and the larger (56 and 54 kDa) form in sperm would be glycosylated at all or some of the five predicted N-glycosylation sites.
The h-tektin-t protein was localized to the principal piece of the human sperm tail, and the discontinuous punctuated staining in sperm flagella may result from a partial masking of h-tektin-t by tubulin. In addition, h-tektin-t was also localized as a broad band in the post-acrosomal region of the sperm head together with two strong spot-like signals which seemed to represent a tangential view of positive signals in the post-acrosomal region. Although the detailed staining pictures showed some differences, these observations were basically similar to those of the earlier report (Wolkowicz et al., 2002). Reports have indicated that various types of tubulin are present in mammalian, including human, sperm head (Draber et al., 1991; Moreno and Schatten, 2000; Smrzka et al., 2000; Peknicova et al., 2001). As tektins are known to associate with tubulins (Stephens, 1994; Norrander et al., 1998), h-tektin-t may be associated with tubulins in the tail as well as in the head of human sperm, and may play some collaborative role with associated tubulins.
Mouse and human-tektin-t proteins had 52 and 53% homology with sea urchin tektin B respectively. The conservation of tektins, especially in the consensus sequence, in rather distantly related species like the sea urchin and mammals indicates a fundamental role, particularly in stabilizing the three-dimensional form of microtubules. The evolutionary conservation of tektins and their association with tubulins make them important and useful molecular targets for the study of microtubular organization.
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Acknowledgements |
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We are grateful to Professor Kiyotaka Toshimori for valuable discussions. This work was supported by Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists.
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Notes |
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4 To whom correspondence should be addressed. E-mail: nishimun{at}biken.osaka-u.ac.jp
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References |
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Amos, W.B., Amos, L.A. and Linck, R.W. (1986) Studies of tektin filaments from flagellar microtubules by immunoelectron microscopy. J. Cell Sci., (Suppl. 5), 55–68.
Chen, R., Perrone, C.A., Amos, L.A. and Linck, R.W. (1993) Tektin B1 from ciliary microtubules: primary structure as deduced from the cDNA sequence and comparison with tektin A1. J. Cell Sci., 106, 909–918.[Abstract]
Donoghue, A.M., Sonstegard, T.S., King, L.M. Smith, E.J. and Burt, D.W. (1999) Turkey sperm mobility influences paternity in the context of competitive fertilization. Biol. Reprod., 61, 422–427.
Draber, P., Draberova, E. and Viklicky, V. (1991) Immunostaining of human spermatozoa with tubulin domain-specific monoclonal antibodies. Recognition of a unique beta-tubulin epitope in the sperm head. Histochemistry, 95, 519–524.[ISI][Medline]
Froman, D.P., Feltmann, A.J. and McLean, D.J. (1997) Increased fecundity resulting from semen donor selection based upon in vitro sperm motility. Poult. Sci., 76, 73–77.
Gupta, D., Oscarson, S., Raju, T.S., Stanley, P., Toone, E.J. and Brewer, C.F. (1996) A comparison of the fine saccharide-binding specificity of Dioclea grandiflora lectin and concanavalin A. Eur. J. Biochem., 242, 320–326.[ISI][Medline]
Harpar, M.K. (1994) Gamete and Zygote transport. In Knobil, E. and Neill, J.D. (eds) The Physiology of Reproduction, Vol.1, 2nd edn. Raven Press, New York, USA, pp. 123–188.
Iguchi, N., Tanaka, H., Fujii, T., Tamura, K., Kaneko, Y., Nojima, H. and Nishimune, Y. (1999) Molecular cloning of haploid germ cell-specific tektin cDNA and analysis of the protein in mouse testis. FEBS Lett., 456, 315–321.[ISI][Medline]
Jaffe, T. and Oates, D.R. (1997) Genetic aspects of infertility. In Lipshultz, L.I. and Howards, S.S. (eds) Infertility in the Male, 3rd edn. Mosby-Year Book Inc., Missouri, USA, pp. 280–304.
Kobata, A. and Yamashita, K. (1993) Fractionation of oligosaccharides by serial affinity chromatography with use of immobilized lectin columns. In Fukuda, M. and Kobata, A. (eds) Practical Approach—Glycoprotein Analysis. Oxford University Press, Oxford, UK, pp. 103–125.
Kuno, M., Yonezawa, N., Amari, S., Hayashi, M., Ono, Y., Kiss, L., Sonohara, K. and Nakano, M. (2000) The presence of a glycosyl phosphatidylinositol-anchored alpha-mannosidase in boar sperm. IUBMB Life, 49, 485–489.[ISI][Medline]
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.[Medline]
Larsson, M., Norrander, J., Graslund, S., Brundell, E., Linck, R., Stahl, S. and Hoog, C. (2000) The spatial and temporal expression of Tekt1, a mouse tektin C homologue, during spermatogenesis suggest that it is involved in the development of the sperm tail basal body and axoneme. Eur. J. Cell Biol., 79, 718–725.[ISI][Medline]
Linck, R.W. and Langevin, G.L. (1982) Structure and chemical composition of insoluble filamentous components of sperm flagellar microtubules. J. Cell Sci., 58, 1–22.[Abstract]
Linck, R.W. and Stephens, R.E. (1987) Biochemical characterization of tektins from sperm flagellar doublet microtubules. J. Cell Biol., 104, 1069–1075.
Linck, R.W., Albertini, D.F., Kenney, D.M. and Langevin, G.L. (1982) Tektin filaments: chemically unique filaments of sperm flagellar microtubules. Prog. Clin. Biol. Res., 80, 127–132.[Medline]
Linck, R.W., Amos, L.A. and Amos, W.B. (1985) Localization of tektin filaments in microtubules of sea urchin sperm flagella by immunoelectron microscopy. J. Cell Biol., 100, 126–135.
Lopes, C.H., La Falci, V.S., Silva, C.E. and Brandelli, A. (2001) Beta-D-glucuronidase is associated with goat sperm cytoskeleton. J. Exp. Zool., 289, 146–152.[ISI][Medline]
Moreno, R.D. and Schatten, G. (2000) Microtubule configurations and post-translational alpha-tubulin modifications during mammalian spermatogenesis. Cell Motil. Cytoskeleton, 4, 235–246.
Nath, D. and Majumder, G.C. (1999) Maturation-dependent modification of the protein phosphorylation profile of isolated goat sperm plasma membrane. J. Reprod. Fertil., 115, 29–37.[Abstract]
Nojima, D., Linck, R.W. and Egelman, E.H. (1995) At least one of the protofilaments in flagellar microtubules is not composed of tubulin. Curr. Biol., 5, 158–167.[ISI][Medline]
Norrander, J.M., Perrone, C.A., Amos, L.A. and Linck, R.W. (1996) Structural comparison of tektins and evidence for their determination of complex spacings in flagellar microtubules. J. Mol. Biol., 257, 385–397.[ISI][Medline]
Norrander, J., Larsson, M., Stahl, S., Hoog, C. and Linck, R. (1998) Expression of ciliary tektins in brain and sensory development. J. Neurosci., 18, 8912–8918.
Norrander, J.M., deCathelineau, A.M., Brown, J.A., Porter, M.E. and Linck, R.W. (2000) The Rib43a protein is associated with forming the specialized protofilament ribbons of flagellar microtubules in Chlamydomonas. Mol. Biol. Cell, 11, 201–215.
Ogata, S., Muramatsu, T. and Kobata, A. (1975) Fractionation of glycopeptides by affinity column chromatography on concanavalin A-sepharose. J. Biochem. (Tokyo), 78, 687–696.
Olesen, C., Hansen, C., Bendsen, E., Byskov, A.G., Schwinger, E., Lopez-Pajares, I., Jensen, P.K., Kristoffersson, U., Schubert, R. and Van Assche, E. et al. (2001) Identification of human candidate genes for male infertility by digital differential display. Mol. Hum. Reprod., 7, 11–20.
Peknicova, J., Kubatova, A., Sulimenko, V., Draberova, E., Viklicky, V., Hozak, P. and Draber, P. (2001) Differential subcellular distribution of tubulin epitopes in boar spermatozoa: recognition of class III beta-tubulin epitope in sperm tail. Biol. Reprod., 65, 672–679.
Pirner, M.A. and Linck, R.W. (1994) Tektins are heterodimeric polymers in flagellar microtubules with axial periodicities matching the tubulin lattice. J. Biol. Chem., 269, 31800–31806.
Russell, L.D., Ettlin, R.A., Sinha, H.A.P. and Clegg, E.D. (1990) Mammalian spermatogenesis. In Russell, L.D., Ettlin, R.A., Sinha, H.A.P. and Clegg, E.D. (eds) Histological and Histopathological Evaluation of the Testis. Cache River Press, Florida, USA, pp. 1–40.
Sanger, F., Nicklen, S. and Coulson, A.R. (1977) DNA sequencing with chain-terminating inhibitors. Proc. Natl Acad. Sci. USA, 74, 5463–5467.
Smrzka, O.W., Delgehyr, N. and Bornens, M. (2000) Tissue-specific expression and subcellular localisation of mammalian delta-tubulin. Curr. Biol., 10, 413–416.[ISI][Medline]
Steffen, W. and Linck, R.W. (1988) Evidence for tektins in centrioles and axonemal microtubules. Proc. Natl Acad. Sci. USA, 85, 2643–2647.
Stephens, R.E. (1994) Tubulin and tektin in sea urchin embryonic cilia: pathways of protein incorporation during turnover and regeneration. J. Cell Sci., 107, 683–692.[Abstract]
Tanaka, H., Ikawa, M., Tsuchida, J., Nozaki, M., Suzuki, M., Fujiwara, T., Okabe, M. and Nishimune, Y. (1997) Cloning and characterization of the human Calmegin gene encoding putative testis-specific chaperone. Gene, 204, 159–163.[ISI][Medline]
Tulsiani, D.R., Orgebin-Crist, M.C. and Skudlarek, M.D. (1998) Role of luminal fluid glycosyltransferases and glycosidases in the modification of rat sperm plasma membrane glycoproteins during epididymal maturation. J. Reprod. Fertil. (Suppl. 53), 85–97.
Watanabe, T.K., Kawai, A., Fujiwara, T., Maekawa, H., Hirai, Y., Nakamura, Y. and Takahashi, E. (1996) Molecular cloning of UBE2G, encoding a human skeletal muscle-specific ubiquitin-conjugating enzyme homologous to UBC7 of C. elegans.Cytogenet. Cell Genet., 74, 146–148.[ISI][Medline]
Wolkowicz, M.J., Naaby-Hansen, S., Gamble, A.R., Reddi, P.P., Flickinger, C.J. and Herr, J.C. (2001) Tektin B1 demonstrates flagellar localization in human sperm. Biol. Reprod., 66, 241–250.
Submitted on November 2, 2001; accepted on March 1, 2002.
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