+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

Sequence-specific interaction with the viral AL1 protein identifies a geminivirus DNA replication origin

Sequence-specific interaction with the viral AL1 protein identifies a geminivirus DNA replication origin

Plant Cell 4(7): 799-809

The bipartite geminiviruses such as tomato golden mosaic virus (TGMV) and squash leaf curl virus (SqLCV) have two single-stranded circular genomic DNAs, the A and B components, thought to be replicated from double-stranded circular DNA intermediates. Although it has been presumed that the origin sequences for viral replication are located in the highly conserved 200-nucleotide common region (CR) present in both genomic components and that the viral-encoded AL1 protein interacts with these sequences to effect replication, there has been no evidence that this is in fact so. We have investigated these questions, demonstrating selectivity and sequence specificity in this protein-DNA interaction. Simple component switching between the DNAs of TGMV and SqLCV and analysis of replication in leaf discs showed that whereas the A components of both TGMV and SqLCV promote their own replication and that of their cognate B component, neither replicates the noncognate B component. Furthermore, using an in vivo functional replication assay, we found that cloned viral CR sequences function as a replication origin and direct the replication of nonviral sequences in the presence of AL1, with both circular single-stranded and double-stranded DNA being synthesized. Finally, by the creation of chimeric viral CRs and specific subfragments of the viral CR, we demonstrated sequence-specific recognition of the replication origin by the AL1 protein, thereby localizing the origin to an approximately 90-nucleotide segment in the AL1 proximal side of the CR that includes the conserved geminiviral stem-loop structure and approximately 60 nucleotides of 5' upstream sequence. By deletional analysis, we further demonstrated that the conserved stem-loop structure is essential for replication. These studies identify the functional viral origin of replication within the CR, demonstrating that sequence-specific recognition of this origin by the AL1 protein is required for replication.

(PDF same-day service: $19.90)

Accession: 002219106

Download citation: RISBibTeXText

PMID: 1392596

DOI: 10.1105/tpc.4.7.799

Related references

Interaction between a geminivirus replication protein and origin DNA is essential for viral replication. Journal of Biological Chemistry 269(11): 8459-8465, 1994

Dual interaction of plant PCNA with geminivirus replication accessory protein (REn) and viral replication protein (Rep). Virology 312(2): 381-394, 2003

Dual interaction of plant PCNA with geminivirus replication accessory protein and viral replication protein. Virology 312(2): 381-394, August 1, 2003

Dual interaction of a geminivirus replication accessory factor with a viral replication protein and a plant cell cycle regulator. Virology 279(2): 570-576, 2001

Sequence-specific interaction between the replication initiator protein of plasmid pT181 and its origin of replication. Proceedings of the National Academy of Sciences of the United States of America 83(15): 5484-5488, 1986

Insights into the functional characteristics of geminivirus rolling-circle replication initiator protein and its interaction with host factors affecting viral DNA replication. Archives of Virology 160(2): 375-387, 2015

Interaction of geminivirus Rep protein with replication factor C and its potential role during geminivirus DNA replication. Virology 302(1): 83-94, 2002

Interaction between geminivirus replication protein and the SUMO-conjugating enzyme is required for viral infection. Journal of Virology 85(19): 9789-9800, 2011

A geminivirus replication protein is a sequence-specific DNA binding protein. Plant Cell 4(5): 597-608, 1992

The interaction of herpes simplex type 1 virus origin-binding protein (UL9 protein) with Box I, the high affinity element of the viral origin of DNA replication. Journal of Biological Chemistry 274(26): 18613-7, 1999

Peptide aptamers that bind to a geminivirus replication protein interfere with viral replication in plant cells. Journal of Virology 80(12): 5841-5853, 2006

The origin binding protein of herpes simplex virus type 1 and its interaction with a viral origin of replication. Journal of Cellular Biochemistry Supplement (16 PART B): 105, 1992

The replication inhibition activity of the WT1 tumor suppressor protein resides in its N-terminal 298 amino acid region, and does not require specific binding of the protein to the replication origin sequence. International Journal of Oncology 13(6): 1275-1280, 1998