Expression analysis of the human testis-specific serine/threonine kinase (TSSK) homologues. A TSSK member is present in the equatorial segment of human sperm

Zhonglin Hao, Kula N. Jha, Young Hwan Kim, Soumya Vemuganti, V. Anne Westbrook, Olga Chertihin, Karin Markgraf, Charles J. Flickinger, Michael Coppola, John C. Herr, Pablo E. Visconti

Research output: Contribution to journalArticle

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Abstract

Two members of the human testis-specific serine/threonine (Ser/Thr) kinase family, TSSK 1 and TSSK 2, were cloned and sequenced from a human testis adaptor-ligated cDNA library using a PCR strategy. Within the cDNA, open reading frames (ORF) were defined encoding proteins of 367 and 358 amino acids respectively, as well as conserved kinase domains typical of the superfamily of Ser/Thr kinases. Both genes were intronless and mapped to chromosomes 5 and 22 respectively. The human and mouse homologues of TSSK 1 and TSSK 2, together with TSSK 3 and SSTK/FKSG82, constitute a kinase subfamily closely related to the calmodulin kinases and SNF/nim 1 kinase subfamilies. Similar to the mouse, tissue expression by northern and dot blot analysis revealed that human TSSK 1 and 2 messages are expressed exclusively in the testis. However, mRNA for these kinases can be detected in other tissues using real-time PCR. In addition, TSKS, the human homologue of a putative substrate of TSSK 1 and 2, was cloned. TSKS had an ORF of 592 amino acids and was also expressed exclusively in the testis as demonstrated by northern and dot blot analyses; however, lower levels of expression in other tissues were detected using real-time PCR. Human TSSK 2 and TSKS interacted in a yeast two-hybrid system and also co-immunoprecipitated after in vitro translation. TSSK 2 expressed in yeast and bacteria was able to autophosphorylate and also phosphorylated recombinant TSKS in vitro. Antibodies against recombinant TSSK 2 demonstrated that a member of the TSSK family was present in human testis and localized to the equatorial segment of ejaculated human sperm. In contrast, TSKS was only found in the testis. The finding of a TSSK family member in mature sperm suggests that this family of kinases might play a role in sperm function.

Original languageEnglish (US)
Pages (from-to)433-444
Number of pages12
JournalMolecular Human Reproduction
Volume10
Issue number6
DOIs
StatePublished - Jun 1 2004

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Protein-Serine-Threonine Kinases
Spermatozoa
Testis
Phosphotransferases
Northern Blotting
Open Reading Frames
Real-Time Polymerase Chain Reaction
Amino Acids
Chromosomes, Human, Pair 22
Calcium-Calmodulin-Dependent Protein Kinases
Chromosomes, Human, Pair 5
Two-Hybrid System Techniques
Gene Library

Keywords

  • Serine
  • Spermatogenesis
  • TSKS
  • TSSK
  • Testis-specific
  • Threonine kinase

ASJC Scopus subject areas

  • Reproductive Medicine
  • Embryology
  • Molecular Biology
  • Genetics
  • Obstetrics and Gynecology
  • Developmental Biology
  • Cell Biology

Cite this

Expression analysis of the human testis-specific serine/threonine kinase (TSSK) homologues. A TSSK member is present in the equatorial segment of human sperm. / Hao, Zhonglin; Jha, Kula N.; Kim, Young Hwan; Vemuganti, Soumya; Westbrook, V. Anne; Chertihin, Olga; Markgraf, Karin; Flickinger, Charles J.; Coppola, Michael; Herr, John C.; Visconti, Pablo E.

In: Molecular Human Reproduction, Vol. 10, No. 6, 01.06.2004, p. 433-444.

Research output: Contribution to journalArticle

Hao, Z, Jha, KN, Kim, YH, Vemuganti, S, Westbrook, VA, Chertihin, O, Markgraf, K, Flickinger, CJ, Coppola, M, Herr, JC & Visconti, PE 2004, 'Expression analysis of the human testis-specific serine/threonine kinase (TSSK) homologues. A TSSK member is present in the equatorial segment of human sperm', Molecular Human Reproduction, vol. 10, no. 6, pp. 433-444. https://doi.org/10.1093/molehr/gah052
Hao, Zhonglin ; Jha, Kula N. ; Kim, Young Hwan ; Vemuganti, Soumya ; Westbrook, V. Anne ; Chertihin, Olga ; Markgraf, Karin ; Flickinger, Charles J. ; Coppola, Michael ; Herr, John C. ; Visconti, Pablo E. / Expression analysis of the human testis-specific serine/threonine kinase (TSSK) homologues. A TSSK member is present in the equatorial segment of human sperm. In: Molecular Human Reproduction. 2004 ; Vol. 10, No. 6. pp. 433-444.
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abstract = "Two members of the human testis-specific serine/threonine (Ser/Thr) kinase family, TSSK 1 and TSSK 2, were cloned and sequenced from a human testis adaptor-ligated cDNA library using a PCR strategy. Within the cDNA, open reading frames (ORF) were defined encoding proteins of 367 and 358 amino acids respectively, as well as conserved kinase domains typical of the superfamily of Ser/Thr kinases. Both genes were intronless and mapped to chromosomes 5 and 22 respectively. The human and mouse homologues of TSSK 1 and TSSK 2, together with TSSK 3 and SSTK/FKSG82, constitute a kinase subfamily closely related to the calmodulin kinases and SNF/nim 1 kinase subfamilies. Similar to the mouse, tissue expression by northern and dot blot analysis revealed that human TSSK 1 and 2 messages are expressed exclusively in the testis. However, mRNA for these kinases can be detected in other tissues using real-time PCR. In addition, TSKS, the human homologue of a putative substrate of TSSK 1 and 2, was cloned. TSKS had an ORF of 592 amino acids and was also expressed exclusively in the testis as demonstrated by northern and dot blot analyses; however, lower levels of expression in other tissues were detected using real-time PCR. Human TSSK 2 and TSKS interacted in a yeast two-hybrid system and also co-immunoprecipitated after in vitro translation. TSSK 2 expressed in yeast and bacteria was able to autophosphorylate and also phosphorylated recombinant TSKS in vitro. Antibodies against recombinant TSSK 2 demonstrated that a member of the TSSK family was present in human testis and localized to the equatorial segment of ejaculated human sperm. In contrast, TSKS was only found in the testis. The finding of a TSSK family member in mature sperm suggests that this family of kinases might play a role in sperm function.",
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AU - Westbrook, V. Anne

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AU - Markgraf, Karin

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