Accession Number:

ADA386576

Title:

Cloning and Biochemical Analysis of the Ataxia-Telangiectasia (A-T) Gene in Xenopus Laevis

Descriptive Note:

Annual rept. 1 Sep 1998-31 Aug 1999

Corporate Author:

COLUMBIA UNIV NEW YORK

Personal Author(s):

Report Date:

1999-09-01

Pagination or Media Count:

43.0

Abstract:

ATM, the gene product mutated in Ataxia Telangiectasia A-T encodes a 35O-kDa protein involved in the regulation of several cellular responses to DNA breaks. We used a degenerate PCR-based strategy to isolate a partial clone of X-ATM, the Xenopus homologue of human ATM. Sequence analysis confirmed that the clone was most closely related to human ATM. Xenopus ATM protein X-ATM is 85 identical to human ATM within the kinase domain and 71 identical over the carboxyl-terminal half of the protein. Polyclonal antibodies raised against recombinant X-ATM are highly specific for the ATM protein and recognize a single polypeptide of 37O-kDa in oocytes, embryos, egg extracts and a Xenopus cell line. We found that X-ATM was expressed maternally in eggs and as early as stage II pre-vitellogenic oocytes, and the protein and mRNA were present at relatively constant levels throughout development. Subcellular fractionation showed that the protein was nuclear in both the female and male germ lines. The level of X-ATM protein did not change throughout the meiotic divisions or the synchronous mitotic cycles of cleavage stage embryos. In addition, we did not observe any change in the level or mobility of X-ATM protein following gamma-irradiation of embryos. Finally, we also demonstrated that X-ATM was present in a high molecular weight complex of approximately 500 kDa containing the X-ATM protein and other, as yet unidentified components. SUBJECT TERMS IS. NUMBER OF PAGES Cancer

Subject Categories:

  • Biochemistry
  • Genetic Engineering and Molecular Biology
  • Medicine and Medical Research

Distribution Statement:

APPROVED FOR PUBLIC RELEASE