Date

June 1997

Document Type

Thesis

Degree Name

M.S.

Department

Dept. of Biochemistry and Molecular Biology

Institution

Oregon Graduate Institute of Science & Technology

Abstract

Neurospora crassa arg-2 encodes the small subunit of Arg-specific carbamoyl phosphate synthetase and is negatively regulated by arginine. This regulation is mediated by a 24-codon upstream open reading frame (uORF). The sequence of this uORF is critical for Arg-specific regulation. Six mutated templates were used to examine which residues of the uORF are important for this regulation. Mutations were created using megaprimer PCR and a luciferase gene was used as a reporter in the in vitro translation studies. Mutations of Asp 12, Asp 16, and Ser 10 eliminate Arg-specific regulation. Leaky scanning is thought to be involved, and a hypothetical ribosome stalling model that mediates Arg-specific attenuation of translation is proposed.

Identifier

doi:10.6083/M45B00CR

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