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DNA binding exerted by a bacterial gene regulator with an extensive coiled-coil domain
Karolinska Institutet.ORCID iD: 0000-0002-3049-967X
1996 (English)In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 271, no 29, 12626-12631 p.Article in journal (Refereed) PublishedText
Abstract [en]

Although quite common in the eukaryotic cell, bacterial proteins with an extensive coiled-coil domain are still relatively rare. One of the few thus far documented examples, TlpA from Salmonella typhimurium, is characterized by a remarkably long (250 amino acids) alpha-helical coiled-coil domain. Herein, we demonstrate that TlpA is a novel, sequence-specific DNA-binding protein. Several tlpA deletion mutants have been constructed, and their corresponding protein products were purified and tested for DNA binding. Two of the mutant proteins were shown to be deficient in DNA binding. Both mutants were analyzed by circular dichroism and electron microscopy, supporting the notion that mutant proteins were largely intact despite lacking the amino acid residues necessary for DNA binding. In vivo studies with transcriptional tlpA-lacZ fusions demonstrated that TlpA acts as a repressor. Using the repressor phenotype as a readout, the chain exchange previously described in vitro could also be confirmed in vivo. We believe the coiled-coil domain acts not only as a dimerization interface but could also serve a role as a flexible modulator of the protein-DNA interaction.

Place, publisher, year, edition, pages
1996. Vol. 271, no 29, 12626-12631 p.
Keyword [en]
design, leucine zipper, protein, transcription factors, virulence plasmid
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:sh:diva-29020DOI: 10.1074/jbc.271.21.12626ISI: A1996UL66000076PubMedID: 8647874ScopusID: 2-s2.0-17544362542OAI: oai:DiVA.org:sh-29020DiVA: diva2:892163
Note

Correction in: Journal of Biological Chemistry, Volume: 271, Issue: 29, Pages: 17592-17592

From Duplicate 1 ( DNA binding exerted by a bacterial gene regulator with an extensive coiled-coil domain - Hurme, R; Berndt, K D; Namork, E; Rhen, M ) Times Cited: 11 Article English Cited References Count: 31 Ul660 From Duplicate 2 ( DNA binding exerted by a bacterial gene regulator with an extensive coiled-coil domain (vol 271, pg 12626, 1996) - Hurme, R; Berndt, K D; Namork, E; Rhen, M ) Times Cited: 0 Correction, Addition English Cited References Count: 1 Ux943

Available from: 2016-01-08 Created: 2016-01-07 Last updated: 2016-01-11Bibliographically approved

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