Display Full Text | Download Full Text
4.48 MB PDF file

Degree

Doctor of Philosophy, Case Western Reserve University, Biochemistry, 1994.

Abstract

Roles of RNA structures in the 5′ non translated region of Rous Sarcoma Virus (RSV) in the initiation of reverse transcription were evaluated. Besides the previously demonstrated U5-leader stem, reverse transcription of RSV was found to require the U5-IR stem. Disruption of the latter by substitution mutations could be compensated by secondary mutations that restored the structure of the stem but not its sequence. Additionally a role for U5 RNA sequences in base-pairing to the TψC loop sequences in primer tRNA Trp was established. This was examined in vivo and in vitro using a substitution mutation that abrogates the U5-TψC interaction. In vitro this mutation could be rescued by using a primer that contained sequences complementary to the substitution. Further studies revealed that these three regions of duplex RNA structure must be maintained at a specific spatial orientation with respect to each other. Disruption of "spacer" sequences between the duplex regions by insertion and deletion mutations caused defects in initiation. In contrast, substitution mutations in these non base-paired spacer sequences had no effect.

Keywords

RNA structures; Rous sarcoma; virus reverse transcription

Advisor

Jonathan P. Leis

Pages

187p.

Document number: case1057948961
Permalink: http://rave.ohiolink.edu/etdc/view?acc_num=case1057948961

This ETD has been downloaded 264 times (through March 2013)