UCL logo

UCL Discovery

UCL home » Library Services » Electronic resources » UCL Discovery

Investigation of TSGA10 gene expression, localization and protein interaction in human and mouse spermatogenesis.

Behnam, B.; (2005) Investigation of TSGA10 gene expression, localization and protein interaction in human and mouse spermatogenesis. Doctoral thesis , University of London. Green open access

[img] Text
U592635.pdf

Download (30MB)

Abstract

Isolation of the testis-specific human gene TSGA10 was reported in 2001 (Modarressi et al.). In this project, I have carried on the work by further characterising the gene and its product by cloning and sequencing the mouse homologue (Tsga10) (GenBank Accession no. AF5300501). Tsga10 was localised by FISH to mouse chromosome 1 band B, and RNA in situ hybridization and RT-PCR experiments showed that Tsga10 transcripts were developmentally regulated and expressed in pachytene cells of the testis. It was also expressed in some other actively dividing cells such as foetal tissues, primary tumors and astrocytes. Interestingly, the program Pfam (which predicts protein structural motifs) suggests that the Tsga10 protein contains a 'myosin like' coiled-coil domain. A Green fluorescent protein (GFP)-Tsgal10 fusion gene was constructed and cell transfection resulted in the formation of short thick protein filaments. Antibodies raised individually against the N-terminus and C-terminus parts of the Tsga10 protein were used to localise Tsga10 protein expression by immunohistochemistry. In collaboration with Hossein Modarressi and Frans Van der Hoorn, using immuno-blotting and staining techniques, I discovered that Tsga10 encodes a 82 kD precursor protein in spermatids which is incorporated into sperm tails. Upon passage of sperm into the epididymis this is processed to a 27 kD protein which is associated with the fibrous sheath, a major tail structure. Using Tsga10 as bait in a yeast two-hybrid experiment, citestis cDNA library was screened for proteins that interacted with Tsga10. Three candidate interacting proteins were found; Odf2, a previously described protein of the fibrous sheath, the rat homologue of FU32880, (a hypothetical human protein whose gene is expressed predominantly in spermatocytes, spermatids and mature sperm cells), and the mitochondrial protein cytochrome cl. Using immunocytochemistry I then localised FLJ32880 (newly named as TSGA10 interacting protein, TSGA10IP) in the sperm tail as another fibrous sheath protein. These results support the theory that Tsga10 has a role to play in the fibrous sheath of the sperm tail, and also suggest a possible functional involvement with a mitochondrial protein. Antibody was raised against the mouse homologue of TSGA10IP for further study. I suggest that Tsga10 plays multiple roles, one in sperm tail formation and possibly a second role in cell cycle regulation.

Type: Thesis (Doctoral)
Title: Investigation of TSGA10 gene expression, localization and protein interaction in human and mouse spermatogenesis.
Identifier: PQ ETD:592635
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by Proquest
UCL classification: UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Genetics, Evolution and Environment
URI: http://discovery.ucl.ac.uk/id/eprint/1445315
Downloads since deposit
199Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item