6QW2

The Transcriptional Regulator PrfA-A218G mutant from Listeria Monocytogenes

6QW2 の概要

• エントリーDOI: 10.2210/pdb6qw2/pdb
• 分子名称: Listeriolysin positive regulatory factor A, SODIUM ION, ISOPROPYL ALCOHOL, ... (4 entities in total)
• 機能のキーワード: transcription regulator, activation, listeria monocytogenes, transcription, virulence factor, dna binding protein
• 由来する生物種: Listeria monocytogenes
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 55173.35

構造登録者

Hall, M.,Grundstrom, C.,Hansen, S.,Brannstrom, K.,Johansson, J.,Sauer-Eriksson, A.E. (登録日: 2019-03-05, 公開日: 2020-04-01, 最終更新日: 2024-01-24)

主引用文献

Hansen, S.,Hall, M.,Grundstrom, C.,Brannstrom, K.,Sauer-Eriksson, A.E.,Johansson, J.
A Novel Growth-Based Selection Strategy Identifies New Constitutively Active Variants of the Major Virulence Regulator PrfA in Listeria monocytogenes.
J.Bacteriol., 202:-, 2020

PubMed Abstract: is a Gram-positive pathogen able to cause severe human infections. Its major virulence regulator is the transcriptional activator PrfA, a member of the Crp/Fnr family of transcriptional regulators. To establish a successful infection, the PrfA protein needs to be in an active conformation, either by binding the cognate inducer glutathione (GSH) or by possessing amino acid substitutions rendering the protein constitutively active (PrfA*). By a yet unknown mechanism, phosphotransferase system (PTS) sugars repress the activity of PrfA. We therefore took a transposon-based approach to identify the mechanism by which PTS sugars repress PrfA activity. For this, we screened a transposon mutant bank to identify clones able to grow in the presence of glucose-6-phosphate as the sole carbon source. Surprisingly, most of the isolated transposon mutants also carried amino acid substitutions in PrfA. In transposon-free strains, the PrfA amino acid substitution mutants displayed growth, virulence factor expression, infectivity, and DNA binding, agreeing with previously identified PrfA* mutants. Hence, the initial growth phenotype observed in the isolated clone was due to the amino acid substitution in PrfA and unrelated to the loci inactivated by the transposon mutant. Finally, we provide structural evidence for the existence of an intermediately activated PrfA state, which gives new insights into PrfA protein activation. The Gram-positive bacterium is a human pathogen affecting mainly the elderly, immunocompromised people, and pregnant women. It can lead to meningoencephalitis, septicemia, and abortion. The major virulence regulator in is the PrfA protein, a transcriptional activator. Using a growth-based selection strategy, we identified mutations in the PrfA protein leading to constitutively active virulence factor expression. We provide structural evidence for the existence of an intermediately activated PrfA state, which gives new insights into PrfA protein activation.

PubMed: 32179627
DOI: 10.1128/JB.00115-20

実験手法

X-RAY DIFFRACTION (2.6 Å)