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Transcriptional activation of Agrobacterium tumefaciens virulence gene promoters in Escherichia coli requires the A. tumefaciens RpoA gene, encoding the alpha subunit of RNA polymerase.
J Bacteriol. 1999 Aug; 181(15):4533-9.JB

Abstract

The two-component regulatory system, composed of virA and virG, is indispensable for transcription of virulence genes within Agrobacterium tumefaciens. However, virA and virG are insufficient to activate transcription from virulence gene promoters within Escherichia coli cells, indicating a requirement for additional A. tumefaciens genes. In a search for these additional genes, we have identified the rpoA gene, encoding the alpha subunit of RNA polymerase (RNAP), which confers significant expression of a virB promoter (virBp)::lacZ fusion in E. coli in the presence of an active transcriptional regulator virG gene. We conducted in vitro transcription assays using either reconstituted E. coli RNAP or hybrid RNAP in which the alpha subunit was derived from A. tumefaciens. The two forms of RNAP were equally efficient in transcription from a sigma(70)-dependent E. coli galP1 promoter; however, only the hybrid RNAP was able to transcribe virBp in a virG-dependent manner. In addition, we provide evidence that the alpha subunit from A. tumefaciens, but not from E. coli, is able to interact with the VirG protein. These data suggest that transcription of virulence genes requires specific interaction between VirG and the alpha subunit of A. tumefaciens and that the alpha subunit from E. coli is unable to effectively interact with the VirG protein. This work provides the basis for future studies designed to examine vir gene expression as well as the T-DNA transfer process in E. coli.

Authors+Show Affiliations

Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

10419950

Citation

Lohrke, S M., et al. "Transcriptional Activation of Agrobacterium Tumefaciens Virulence Gene Promoters in Escherichia Coli Requires the A. Tumefaciens RpoA Gene, Encoding the Alpha Subunit of RNA Polymerase." Journal of Bacteriology, vol. 181, no. 15, 1999, pp. 4533-9.
Lohrke SM, Nechaev S, Yang H, et al. Transcriptional activation of Agrobacterium tumefaciens virulence gene promoters in Escherichia coli requires the A. tumefaciens RpoA gene, encoding the alpha subunit of RNA polymerase. J Bacteriol. 1999;181(15):4533-9.
Lohrke, S. M., Nechaev, S., Yang, H., Severinov, K., & Jin, S. J. (1999). Transcriptional activation of Agrobacterium tumefaciens virulence gene promoters in Escherichia coli requires the A. tumefaciens RpoA gene, encoding the alpha subunit of RNA polymerase. Journal of Bacteriology, 181(15), 4533-9.
Lohrke SM, et al. Transcriptional Activation of Agrobacterium Tumefaciens Virulence Gene Promoters in Escherichia Coli Requires the A. Tumefaciens RpoA Gene, Encoding the Alpha Subunit of RNA Polymerase. J Bacteriol. 1999;181(15):4533-9. PubMed PMID: 10419950.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Transcriptional activation of Agrobacterium tumefaciens virulence gene promoters in Escherichia coli requires the A. tumefaciens RpoA gene, encoding the alpha subunit of RNA polymerase. AU - Lohrke,S M, AU - Nechaev,S, AU - Yang,H, AU - Severinov,K, AU - Jin,S J, PY - 1999/7/27/pubmed PY - 1999/7/27/medline PY - 1999/7/27/entrez SP - 4533 EP - 9 JF - Journal of bacteriology JO - J. Bacteriol. VL - 181 IS - 15 N2 - The two-component regulatory system, composed of virA and virG, is indispensable for transcription of virulence genes within Agrobacterium tumefaciens. However, virA and virG are insufficient to activate transcription from virulence gene promoters within Escherichia coli cells, indicating a requirement for additional A. tumefaciens genes. In a search for these additional genes, we have identified the rpoA gene, encoding the alpha subunit of RNA polymerase (RNAP), which confers significant expression of a virB promoter (virBp)::lacZ fusion in E. coli in the presence of an active transcriptional regulator virG gene. We conducted in vitro transcription assays using either reconstituted E. coli RNAP or hybrid RNAP in which the alpha subunit was derived from A. tumefaciens. The two forms of RNAP were equally efficient in transcription from a sigma(70)-dependent E. coli galP1 promoter; however, only the hybrid RNAP was able to transcribe virBp in a virG-dependent manner. In addition, we provide evidence that the alpha subunit from A. tumefaciens, but not from E. coli, is able to interact with the VirG protein. These data suggest that transcription of virulence genes requires specific interaction between VirG and the alpha subunit of A. tumefaciens and that the alpha subunit from E. coli is unable to effectively interact with the VirG protein. This work provides the basis for future studies designed to examine vir gene expression as well as the T-DNA transfer process in E. coli. SN - 0021-9193 UR - https://www.unboundmedicine.com/medline/citation/10419950/Transcriptional_activation_of_Agrobacterium_tumefaciens_virulence_gene_promoters_in_Escherichia_coli_requires_the_A__tumefaciens_RpoA_gene_encoding_the_alpha_subunit_of_RNA_polymerase_ L2 - http://jb.asm.org/cgi/pmidlookup?view=long&pmid=10419950 DB - PRIME DP - Unbound Medicine ER -