Structural and biophysical studies of HIV-1 Rev protein

Rev is a key HIV-1 virus regulatory protein, which exports the unspliced and partially\r\nspliced viral mRNA out of the nucleus, thereby inducing the switch from the early phase\r\nto the late phase of the viral replication cycle. Because it is prone to form filaments at\r\nvery low concentration, the X-ray crystallography or NMR approaches to solving Rev\r\nstructure are very difficult. In my dissertation studies, I have designed several structural\r\nmutants of Rev. Under physiological conditions, I found RevC mutant behaves as a\r\nnatively unfolded monomer in solution. The segmentally disordered character of RevC\r\nwas supported by CD and HSQC studies. Another structural Rev mutant is the Rev loop\r\ndeletion. I used a Gly-Ser-Gly-Ala linker to take the place of the Rev loop and most of\r\nthe Arg-rich domain. This mutant can stay in solution as oligomers in sub-millimolar\r\nconcentration for NMR studies. The HSQC spectra of the Rev loop deletion and the\r\nRevC can be well overlaid except the HSQC of Rev loop deletion which has less peaks\r\nunder the same threshold. It brings a filament model with a thin wall formed by Rev Nterminal\r\ntwo helixes and a flexible Rev C-terminal freely moving inside. The flexible\r\ncharacter of Rev C-terminal is also proved to be true for the Rev wild type in comparing\r\nthe HSQC of Rev wild type and RevC. The HSQC of Rev wild type was obtained from\r\n15N labeled Rev and 132B23 complex. B23 is a major cellular component which binds\r\nRev. I designed an N-terminal core of B23, 132B23, and found that it binds with Rev and\r\nforms a decamer with Rev and 132B23 heterodimer.

Name thesis:degree_name

PhD

Level thesis:degree_level

Doctoral

Grantor

The University of Texas Medical Branch

Year dc:date.issued

2006

Author dc:creator

• Ye Peng

Advisor dc:contributor.advisor

• Robert O. Fox, Ph.D.

Committee members dc:contributor.committeemember

• Vince J. Hilser, Ph.D.
• Susan Marriott, Ph.D.
• Miles W. Cloyd, Ph.D.
• Edmund W. Czerwinski, Ph.D.

• Rev
• HIV-1
• B23

Statement dc:rights

• Copyright © is held by the author. Presentation of this material on the TDL web site by The University of Texas Medical Branch at Galveston was made possible under a limited license grant from the author who has retained all copyrights in the works.

Language dc:language.iso

eng

Dc Identifier Other

etd-03292006-105724

OAI identifier oai:identifier

oai:utmb-ir.tdl.org:2152.3/70