Title

Rapid Evolution of in vivo-Selected Sequences and Structures Replacing 20% of a Subviral RNA

Roles

Student co-authors:
Allison M. Murawski
Johnathan L. Nieves
Holleh F. Tajalli
Tareq Azad
Nina B. Jean-Jacques

Document Type

Article

Publication Date

9-2015

Department

Biology

Language

English

Publication Title

Virology

Abstract

The 356 nt noncoding satellite RNA C (satC) of Turnip crinkle virus (TCV) is composed of 5′ sequences from a second TCV satRNA (satD) and 3′ sequences derived from TCV. SHAPE structure mapping revealed that 76 nt in the poorly-characterized satD-derived region form an extended hairpin (H2). Pools of satC in which H2 was replaced with 76, 38, or 19 random nt were co-inoculated with TCV helper virus onto plants and satC fitness assessed using in vivo functional selection (SELEX). The most functional progeny satCs, including one as fit as wild-type, contained a 38–39 nt H2 region that adopted a hairpin structure and exhibited an increased ratio of dimeric to monomeric molecules. Some progeny of satC with H2 deleted featured a duplication of 38 nt, partially rebuilding the deletion. Therefore, H2 can be replaced by a 38–39 nt hairpin, sufficient for overall structural stability of the 5′ end of satC.

Comments

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DOI

10.1016/j.virol.2015.04.002

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