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Molecular Biotechnology Programme Uppsala University School of Engineering UPTEC X 01 015
Date of issue 2001-03Author
Per Nilsson
Title (English)
Poly(A)-specific ribonuclease; oligomeric structure and cap- binding site
Title (Swedish) Abstract
A Poly(A)-specific exoribonuclease (PARN) has been investigated. The oligomeric structure of PARN was studied by performing cross-linking reactions. Homobifunctional cross-linkers were used and the formed complexes were characterised. PARN was found to be a trimer. One important tool in this study was the development of polyclonal antibodies raised against PARN. The intrinsic property of PARN to bind 5'cap structure of mRNA was further studied. Two PARN mutants were created by site directed mutagenesis. In one mutant Trp-219 was changed to alanine and in the other Glu-455 and Trp-456 were both mutated to alanines. The PARN mutants showed less deadenylating activity and were less bound to the 7-Methyl-GTP-Sepharose.
Keywords:
Deadenylation, mRNA degradation, oligomeric structure, cap Supervisors
Anders Virtanen, Uppsala Univerity
Javier Martinez, Max-Planck-Institut, Göttingen, Germany Examiner
Leif Kirsebom, Uppsala University
Project name Sponsors
Language
English Security
ISSN 1401-2138 Classification
Supplementary bibliographical information Pages
20
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