Tags

Type your tag names separated by a space and hit enter

A key hydrophobic patch identified in an AAA⁺ protein essential for its in trans inhibitory regulation.
J Mol Biol. 2013 Aug 09; 425(15):2656-69.JM

Abstract

Bacterial enhancer binding proteins (bEBPs) are a subclass of the AAA(+) (ATPases Associated with various cellular Activities) protein family. They are responsible for σ(54)-dependent transcription activation during infection and function under many stressful growth conditions. The majority of bEBPs are regulated in their formation of ring-shaped hexameric self-assemblies via an amino-terminal domain through its phosphorylation or ligand binding. In contrast, the Escherichia coli phage shock protein F (PspF) is negatively regulated in trans by phage shock protein A (PspA). Up to six PspA subunits suppress PspF hexamer action. Here, we present biochemical evidence that PspA engages across the side of a PspF hexameric ring. We identify three key binding determinants located in a surface-exposed 'W56 loop' of PspF, which form a tightly packed hydrophobic cluster, the 'YLW' patch. We demonstrate the profound impact of the PspF W56 loop residues on ATP hydrolysis, the σ(54) binding loop 1, and the self-association interface. We infer from single-chain studies that for complete PspF inhibition to occur, more than three PspA subunits need to bind a PspF hexamer with at least two binding to adjacent PspF subunits. By structural modelling, we propose that PspA binds to PspF via its first two helical domains. After PspF binding-induced conformational changes, PspA may then share structural similarities with a bEBP regulatory domain.

Authors+Show Affiliations

Division of Cell and Molecular Biology, Sir Alexander Fleming Building, Imperial College London, Exhibition Road, London SW7 2AZ, UK. nan.zhang@imperial.ac.ukNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

23659791

Citation

Zhang, Nan, et al. "A Key Hydrophobic Patch Identified in an AAA⁺ Protein Essential for Its in Trans Inhibitory Regulation." Journal of Molecular Biology, vol. 425, no. 15, 2013, pp. 2656-69.
Zhang N, Simpson T, Lawton E, et al. A key hydrophobic patch identified in an AAA⁺ protein essential for its in trans inhibitory regulation. J Mol Biol. 2013;425(15):2656-69.
Zhang, N., Simpson, T., Lawton, E., Uzdavinys, P., Joly, N., Burrows, P., & Buck, M. (2013). A key hydrophobic patch identified in an AAA⁺ protein essential for its in trans inhibitory regulation. Journal of Molecular Biology, 425(15), 2656-69. https://doi.org/10.1016/j.jmb.2013.04.024
Zhang N, et al. A Key Hydrophobic Patch Identified in an AAA⁺ Protein Essential for Its in Trans Inhibitory Regulation. J Mol Biol. 2013 Aug 9;425(15):2656-69. PubMed PMID: 23659791.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A key hydrophobic patch identified in an AAA⁺ protein essential for its in trans inhibitory regulation. AU - Zhang,Nan, AU - Simpson,Timothy, AU - Lawton,Edward, AU - Uzdavinys,Povilas, AU - Joly,Nicolas, AU - Burrows,Patricia, AU - Buck,Martin, Y1 - 2013/05/07/ PY - 2013/03/06/received PY - 2013/04/11/revised PY - 2013/04/20/accepted PY - 2013/5/11/entrez PY - 2013/5/11/pubmed PY - 2013/9/10/medline KW - AAA(+) KW - AAA(+) proteins KW - ATPases Associated with various cellular Activities KW - EDTA KW - PDB KW - Protein Data Bank KW - PspA KW - PspF KW - RNA polymerase KW - RNAP KW - WT KW - bEBP KW - bacterial enhancer binding protein KW - enhancer binding protein KW - ethylenediaminetetraacetic acid KW - phage shock protein A KW - phage shock protein F KW - wild type KW - σ54 SP - 2656 EP - 69 JF - Journal of molecular biology JO - J Mol Biol VL - 425 IS - 15 N2 - Bacterial enhancer binding proteins (bEBPs) are a subclass of the AAA(+) (ATPases Associated with various cellular Activities) protein family. They are responsible for σ(54)-dependent transcription activation during infection and function under many stressful growth conditions. The majority of bEBPs are regulated in their formation of ring-shaped hexameric self-assemblies via an amino-terminal domain through its phosphorylation or ligand binding. In contrast, the Escherichia coli phage shock protein F (PspF) is negatively regulated in trans by phage shock protein A (PspA). Up to six PspA subunits suppress PspF hexamer action. Here, we present biochemical evidence that PspA engages across the side of a PspF hexameric ring. We identify three key binding determinants located in a surface-exposed 'W56 loop' of PspF, which form a tightly packed hydrophobic cluster, the 'YLW' patch. We demonstrate the profound impact of the PspF W56 loop residues on ATP hydrolysis, the σ(54) binding loop 1, and the self-association interface. We infer from single-chain studies that for complete PspF inhibition to occur, more than three PspA subunits need to bind a PspF hexamer with at least two binding to adjacent PspF subunits. By structural modelling, we propose that PspA binds to PspF via its first two helical domains. After PspF binding-induced conformational changes, PspA may then share structural similarities with a bEBP regulatory domain. SN - 1089-8638 UR - https://www.unboundmedicine.com/medline/citation/23659791/A_key_hydrophobic_patch_identified_in_an_AAA⁺_protein_essential_for_its_in_trans_inhibitory_regulation_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(13)00272-6 DB - PRIME DP - Unbound Medicine ER -