Abstract
Barley stripe mosaic virus contains seven genes, one of which specifies a 17-kD cysteine-rich protein, gamma b, that is known to affect virulence. To further characterize the role of gamma b in pathogenesis, we mutagenized sequences encoding amino acids within two clusters of cysteine and histidine residues in the cysteine-rich domain and a group of basic amino acids located between the clusters and determined the effects of these mutations on the symptom phenotype in barley. Three single amino acid substitutions in cluster 1 and two amino acid exchanges in the basic region caused bleached symptoms associated with pronounced elevations in accumulation of gamma b protein. In contrast, three single amino acid substitutions in cluster 2 and a mutation in the basic motif resulted in attenuated ("null") symptoms typical of those produced when the gamma b gene is deleted. Tissue infected with these "null" mutants accumulated slightly elevated amounts of the gamma b protein but significantly lower levels of coat protein and the putative movement protein beta b. Genetic complementation tests revealed that cluster 1 mutations are dominant over the wild-type gamma b gene, whereas those in cluster 2 are recessive. These results highlight the pivotal role of gamma b in pathogenesis and suggest that the two cysteine-rich clusters are functionally distinct and that they affect different aspects of disease development.
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- Agranovsky A. A., Karasev A. V., Novikov V. K., Lunina N. A., Loginov S., Tyulkina L. G. Poa semilatent virus, a hordeivirus having no internal polydisperse poly(A) in the 3' non-coding region of the RNA genome. J Gen Virol. 1992 Aug;73(Pt 8):2085–2092. doi: 10.1099/0022-1317-73-8-2085. [DOI] [PubMed] [Google Scholar]
- Atreya C. D., Pirone T. P. Mutational analysis of the helper component-proteinase gene of a potyvirus: effects of amino acid substitutions, deletions, and gene replacement on virulence and aphid transmissibility. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11919–11923. doi: 10.1073/pnas.90.24.11919. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bowles N. E., Damay P., Spahr P. F. Effect of rearrangements and duplications of the Cys-His motifs of Rous sarcoma virus nucleocapsid protein. J Virol. 1993 Feb;67(2):623–631. doi: 10.1128/jvi.67.2.623-631.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Covey S. N. Amino acid sequence homology in gag region of reverse transcribing elements and the coat protein gene of cauliflower mosaic virus. Nucleic Acids Res. 1986 Jan 24;14(2):623–633. doi: 10.1093/nar/14.2.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Donald R. G., Zhou H., Jackson A. O. Serological analysis of barley stripe mosaic virus-encoded proteins in infected barley. Virology. 1993 Aug;195(2):659–668. doi: 10.1006/viro.1993.1417. [DOI] [PubMed] [Google Scholar]
- Dorfman T., Luban J., Goff S. P., Haseltine W. A., Göttlinger H. G. Mapping of functionally important residues of a cysteine-histidine box in the human immunodeficiency virus type 1 nucleocapsid protein. J Virol. 1993 Oct;67(10):6159–6169. doi: 10.1128/jvi.67.10.6159-6169.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fütterer J., Kiss-László Z., Hohn T. Nonlinear ribosome migration on cauliflower mosaic virus 35S RNA. Cell. 1993 May 21;73(4):789–802. doi: 10.1016/0092-8674(93)90257-q. [DOI] [PubMed] [Google Scholar]
- Gorelick R. J., Chabot D. J., Rein A., Henderson L. E., Arthur L. O. The two zinc fingers in the human immunodeficiency virus type 1 nucleocapsid protein are not functionally equivalent. J Virol. 1993 Jul;67(7):4027–4036. doi: 10.1128/jvi.67.7.4027-4036.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gramstat A., Courtpozanis A., Rohde W. The 12 kDa protein of potato virus M displays properties of a nucleic acid-binding regulatory protein. FEBS Lett. 1990 Dec 10;276(1-2):34–38. doi: 10.1016/0014-5793(90)80500-i. [DOI] [PubMed] [Google Scholar]
- Koonin E. V., Boyko V. P., Dolja V. V. Small cysteine-rich proteins of different groups of plant RNA viruses are related to different families of nucleic acid-binding proteins. Virology. 1991 Mar;181(1):395–398. doi: 10.1016/0042-6822(91)90512-a. [DOI] [PubMed] [Google Scholar]
- Koonin E. V., Dolja V. V. Evolution and taxonomy of positive-strand RNA viruses: implications of comparative analysis of amino acid sequences. Crit Rev Biochem Mol Biol. 1993;28(5):375–430. doi: 10.3109/10409239309078440. [DOI] [PubMed] [Google Scholar]
- Kraft R., Tardiff J., Krauter K. S., Leinwand L. A. Using mini-prep plasmid DNA for sequencing double stranded templates with Sequenase. Biotechniques. 1988 Jun;6(6):544-6, 549. [PubMed] [Google Scholar]
- Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Miller E. S., Winter R. B., Campbell K. M., Power S. D., Gold L. Bacteriophage T4 regA protein. Purification of a translational repressor. J Biol Chem. 1985 Oct 25;260(24):13053–13059. [PubMed] [Google Scholar]
- Morozov SYu, Dolja V. V., Atabekov J. G. Probable reassortment of genomic elements among elongated RNA-containing plant viruses. J Mol Evol. 1989 Jul;29(1):52–62. doi: 10.1007/BF02106181. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Niesbach-Klösgen U., Guilley H., Jonard G., Richards K. Immunodetection in vivo of beet necrotic yellow vein virus-encoded proteins. Virology. 1990 Sep;178(1):52–61. doi: 10.1016/0042-6822(90)90378-5. [DOI] [PubMed] [Google Scholar]
- Petty I. T., Donald R. G., Jackson A. O. Multiple genetic determinants of barley stripe mosaic virus influence lesion phenotype on Chenopodium amaranticolor. Virology. 1994 Jan;198(1):218–226. doi: 10.1006/viro.1994.1024. [DOI] [PubMed] [Google Scholar]
- Petty I. T., Edwards M. C., Jackson A. O. Systemic movement of an RNA plant virus determined by a point substitution in a 5' leader sequence. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8894–8897. doi: 10.1073/pnas.87.22.8894. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Petty I. T., French R., Jones R. W., Jackson A. O. Identification of barley stripe mosaic virus genes involved in viral RNA replication and systemic movement. EMBO J. 1990 Nov;9(11):3453–3457. doi: 10.1002/j.1460-2075.1990.tb07553.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Petty I. T., Hunter B. G., Jackson A. O. A novel strategy for one-step cloning of full-length cDNA and its application to the genome of barley stripe mosaic virus. Gene. 1988 Dec 30;74(2):423–432. doi: 10.1016/0378-1119(88)90175-8. [DOI] [PubMed] [Google Scholar]
- Petty I. T., Hunter B. G., Wei N., Jackson A. O. Infectious barley stripe mosaic virus RNA transcribed in vitro from full-length genomic cDNA clones. Virology. 1989 Aug;171(2):342–349. doi: 10.1016/0042-6822(89)90601-6. [DOI] [PubMed] [Google Scholar]
- Petty I. T., Jackson A. O. Mutational analysis of barley stripe mosaic virus RNA beta. Virology. 1990 Dec;179(2):712–718. doi: 10.1016/0042-6822(90)90138-h. [DOI] [PubMed] [Google Scholar]
- Petty I. T., Jackson A. O. Two forms of the major barley stripe mosaic virus nonstructural protein are synthesized in vivo from alternative initiation codons. Virology. 1990 Aug;177(2):829–832. doi: 10.1016/0042-6822(90)90559-a. [DOI] [PubMed] [Google Scholar]
- Robaglia C., Durand-Tardif M., Tronchet M., Boudazin G., Astier-Manifacier S., Casse-Delbart F. Nucleotide sequence of potato virus Y (N Strain) genomic RNA. J Gen Virol. 1989 Apr;70(Pt 4):935–947. doi: 10.1099/0022-1317-70-4-935. [DOI] [PubMed] [Google Scholar]
- Scholthof H. B., Wu F. C., Kiernan J. M., Shepherd R. J. The putative zinc finger of a caulimovirus is essential for infectivity but does not influence gene expression. J Gen Virol. 1993 Apr;74(Pt 4):775–780. doi: 10.1099/0022-1317-74-4-775. [DOI] [PubMed] [Google Scholar]
- Sehnke P. C., Mason A. M., Hood S. J., Lister R. M., Johnson J. E. A "zinc-finger"-type binding domain in tobacco streak virus coat protein. Virology. 1989 Jan;168(1):48–56. doi: 10.1016/0042-6822(89)90402-9. [DOI] [PubMed] [Google Scholar]
- Struhl K. Helix-turn-helix, zinc-finger, and leucine-zipper motifs for eukaryotic transcriptional regulatory proteins. Trends Biochem Sci. 1989 Apr;14(4):137–140. doi: 10.1016/0968-0004(89)90145-X. [DOI] [PubMed] [Google Scholar]
- Ziegler-Graff V., Guilford P. J., Baulcombe D. C. Tobacco rattle virus RNA-1 29K gene product potentiates viral movement and also affects symptom induction in tobacco. Virology. 1991 May;182(1):145–155. doi: 10.1016/0042-6822(91)90658-x. [DOI] [PubMed] [Google Scholar]