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Nucleic Acids Research, 1988, Vol. 16, No. 14 7043-7060
© 1988

Articles

Genetic analysis of the tomato golden mosaic virus II. The product of the AL1 coding sequence is required for replication

J.Scott Elmer, Leslie Brand, Garry Sunter1, William E. Gardiner1, David M. Bisaro1 and Stephen G. Rogers*

Plant Molecular Biology, Biological Sciences, Monsanto Company St Louis, MO 63198 1Department of Molecular Genetics and Ohio State Biotechnology Center, Rightmire Hall, Ohio State University Columbus, OH 43210, USA

*To whom correspondence should be addressed

Received December 31, 1987. Revised May 17, 1988. Accepted May 17, 1988.

Tomato golden mosaic virus (TGMV) belongs to the geminivirus subgroup that is characterized by a split genome consisting of two single-stranded circular DNAs. The TGMV A genome component encodes the virus coat protein as well as all of the functions necessary for viral DNA replication. Analysis of the nucleotide sequence indicates that the TGMV A component has, in addition to the coat protein encoding ORF, four overlapping open reading frames (ORFs) with the potential to encode proteins of greater than 10 kD. We have investigated the functions of these putative proteins in both symptom formation and DNA replication by creating mutations in each of the ORFs. Our results show that the AL4 ORF, which is encoded within the N-terminal region of ORF AL1, is not essential for normal virus infection. In contrast, we find that disruption of the AL3 ORF results in delay and attenuation of symptom formation. We also report that the products of the AL1 and AL2 ORFs are absolutely required for symptom formation. Studies of DNA replication show that only the AL1 open reading frame is essential for viral DNA synthesis. The significance of these results for the development of vectors from the geminiviruses is discussed.


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