Cloning of Carp Septin7 and Its Interaction with sperm specific protein, Msap
Date Issued
2005
Date
2005
Author(s)
Lee, Yi-Tsun
DOI
zh-TW
Abstract
Septin is a highly conserved group of cytoskeleton protein that have been found in almost all eukaryotes but not in protozoa or plants. The septin genes were originally identified in Saccharomyces cerevisiae, and have an important role during cytokinesis. The budding yeast mutants that exhibited defects in septins will produce abnormal morphology during the budding process. Recent researches implicate that different Septins have diverse cellular roles including cell polarity, programmed cell death, cell cycle regulation and cell morphogenesis. The septin family of genes possess three common structures: an N-terminal basic domain, GTPase domain and a C-terminal coiled-coil domain. In mammal cells, the function of the N-terminal basic region is possibly responsible for the interaction with phosphoinositides. Some data point that the C-terminal coiled-coil domain is also responsible for intermolecular interaction upon Septin complex formation.
Previous studies about the spermatogenesis associated gene, msap, of our laboratory by using two dimensional electrophoresis discovered the possible interaction between Septin7 and Msap. The studies also revealed that Septins are localized in the basal body of carp sperm, which is consistent with other research. In this study, the septin7 cDNA is cloned by SMART RACE system and two variants with different length are obtained. By using PCR, we found that both two transcripts exist in all kind of tissue, but the relative expression is different. The result of immunohistochemistry revealed that the Septin7 is ubiquitously expressed present in all testicular tissue. Western blot by Septin7 antibody showed the Septin7 is expressed as 50 kDa as a major form in all kind of tissue, althouth enriched in testis. To confirm the interaction between Septin7 and Msap, pull down and co-immunoprecipitation were used. The results favored the possibility that Septin7 interacts with Msap.
Subjects
鯉魚
精子
Septin7
Msap
Type
other
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