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THE PLANT CELL, Vol 6, Issue 8 1157-1170, Copyright © 1994 by American Society of Plant Biologists
A DNA Sequence Required for Geminivirus Replication Also Mediates Transcriptional Regulation
P. A. Eagle, B. M. Orozco and L. Hanley-Bowdoin
Department of Biochemistry, North Carolina State University, Raleigh, North Carolina 27695-7622
Tomato golden mosaic virus (TGMV), a member of the geminivirus family,
requires a single virus-encoded protein for DNA replication. We show that
the TGMV replication protein, AL1, also acts during transcription to
specifically repress the activity of its promoter. An earlier study
established that AL1 binds to a 13-bp sequence (5[prime]-GGTAGTAAGGTAG)
that is essential for activity of the TGMV replication origin. Analysis of
AL1 binding site mutants in transient expression assays demonstrated that
the same site, which is located between the transcription start site and
TATA box in the AL1 promoter, also mediates transcriptional repression.
These experiments revealed that the repeated motifs in the AL1 binding site
contribute differentially to repression, as has been observed previously
for AL1-DNA binding and viral replication. Introduction of the AL1 binding
site into the 35S promoter of the cauliflower mosaic virus was sufficient
to confer AL1-mediated repression to the heterologous promoter. Analysis of
a truncated AL1 promoter and of mutant AL1 proteins showed that repression
does not require a replication-competent template or a
replication-competent AL1 protein. Transient expression studies using two
different Nicotiana cell lines revealed that, although the two lines
replicate plasmids containing the TGMV origin similarly, they support very
different levels of AL1-mediated repression. These results suggest that
geminivirus transcriptional repression and replication may be independent
processes.
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