Nucleic Acids Research, Vol 26, Issue 10 2426-2432, Copyright © 1998 by Oxford University Press
CD Carpenter and AE Simon
In vivo genetic selection was used to study the sequences and structures
required for accumulation of subviral sat-RNA C associated with turnip
crinkle virus (TCV). This technique is advantageous over site-specific
mutagenesis by allowing side-by-side selection from numerous sequence
possibilities as well as sequence evolution. A 22 base hairpin and 6 base
single-stranded tail located at the 3'-terminus of sat-RNA C were
previously identified as the promoter for minus strand synthesis.
Approximately 50% of plants co-inoculated with TCV and sat-RNA C containing
randomized sequence in place of the 22 base hairpin accumulated sat-RNA in
uninoculated leaves. The 22 base region differed in sat-RNA accumulating in
all infected plants, but nearly all were predicted to fold into a hairpin
structure that maintained the 6 base tail as a single-stranded sequence.
Two additional rounds of sat- RNA amplification led to four sequence family
'winners', with three families containing multiple variants, indicating
that evolution of these sequences was occurring in plants. Three of the
four sequence family winners had the same 3 bp at the base of the stem as
wild-type sat-RNA C. Two of the winners shared 15 of 22 identical bases,
including the entire stem region and extending two bases into the loop.
These results demonstrate the utility of the in vivo selection approach by
showing that both sequence and structure contribute to a more active 3'-end
region for accumulation of sat-RNA C.
ARTICLES
Analysis of sequences and predicted structures required for viral satellite RNA accumulation by in vivo genetic selection
Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, MA 01003, USA.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  J. A. Khan, Q. Wang, R. D. Sjolund, A. Schulz, and G. A. Thompson An Early Nodulin-Like Protein Accumulates in the Sieve Element Plasma Membrane of Arabidopsis Plant Physiology, April 1, 2007; 143(4): 1576 - 1589. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Li and S.-M. Wong Host-dependent effects of the 3' untranslated region of turnip crinkle virus RNA on accumulation in Hibiscus and Arabidopsis J. Gen. Virol., February 1, 2007; 88(2): 680 - 687. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Rico, P. Ivars, S. F. Elena, and C. Hernandez Insights into the selective pressures restricting pelargonium flower break virus genome variability: evidence for host adaptation. J. Virol., August 1, 2006; 80(16): 8124 - 8132. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. ZHANG, J. ZHANG, A. T. GEORGE, T. BAUMSTARK, and A. E. SIMON Conformational changes involved in initiation of minus-strand synthesis of a virus-associated RNA RNA, January 1, 2006; 12(1): 147 - 162. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L.-H. Guo, Y.-H. Cao, D.-W. Li, S.-N. Niu, Z.-N. Cai, C.-G. Han, Y.-F. Zhai, and J.-L. Yu Analysis of Nucleotide Sequences and Multimeric Forms of a Novel Satellite RNA Associated with Beet Black Scorch Virus J. Virol., March 15, 2005; 79(6): 3664 - 3674. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Zhang, J. Zhang, and A. E. Simon Repression and Derepression of Minus-Strand Synthesis in a Plus-Strand RNA Virus Replicon J. Virol., July 15, 2004; 78(14): 7619 - 7633. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. McCormack and A. E. Simon Biased Hypermutagenesis Associated with Mutations in an Untranslated Hairpin of an RNA Virus J. Virol., July 15, 2004; 78(14): 7813 - 7817. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Man and B. L. Epel Characterization of regulatory elements within the coat protein (CP) coding region of Tobacco mosaic virus affecting subgenomic transcription and green fluorescent protein expression from the CP subgenomic RNA promoter J. Gen. Virol., June 1, 2004; 85(6): 1727 - 1738. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. I. Song and W. A. Miller cis and trans Requirements for Rolling Circle Replication of a Satellite RNA J. Virol., March 15, 2004; 78(6): 3072 - 3082. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Sun and A. E. Simon Fitness of a Turnip Crinkle Virus Satellite RNA Correlates with a Sequence-Nonspecific Hairpin and Flanking Sequences That Enhance Replication and Repress the Accumulation of Virions J. Virol., July 15, 2003; 77(14): 7880 - 7889. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C.-Y. Huang, Y.-L. Huang, M. Meng, Y.-H. Hsu, and C.-H. Tsai Sequences at the 3' Untranslated Region of Bamboo Mosaic Potexvirus RNA Interact with the Viral RNA-Dependent RNA Polymerase J. Virol., March 15, 2001; 75(6): 2818 - 2824. [Abstract] [Full Text]
|
|
|
|
|
|
J. Wang and A. E. Simon 3'-End Stem-Loops of the Subviral RNAs Associated with Turnip Crinkle Virus Are Involved in Symptom Modulation and Coat Protein Binding J. Virol., July 15, 2000; 74(14): 6528 - 6537. [Abstract] [Full Text]
|
|