CheZ Phosphatase Localizes to Chemoreceptor Patches via CheA-Short
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| Published in: | Journal of Bacteriology Vol. 185; no. 7; pp. 2354 - 2361 |
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| AbstractList | We have investigated the conditions required for polar localization of the CheZ phosphatase by using a CheZ-green fluorescent protein fusion protein that, when expressed from a single gene in the chromosome, restored chemotaxis to a DeltacheZ strain. Localization was observed in wild-type, DeltacheZ, DeltacheYZ, and DeltacheRB cells but not in cells with cheA, cheW, or all chemoreceptor genes except aer deleted. Cells making only CheA-short (CheA(S)) or CheA lacking the P2 domain also retained normal localization, whereas cells producing only CheA-long or CheA missing the P1 and P2 domains did not. We conclude that CheZ localization requires the truncated C-terminal portion of the P1 domain present in CheA(S). Missense mutations targeting residues 83 through 120 of CheZ also abolished localization. Two of these mutations do not disrupt chemotaxis, indicating that they specifically prevent interaction with CheA(S) while leaving other activities of CheZ intact. We have investigated the conditions required for polar localization of the CheZ phosphatase by using a CheZ-green fluorescent protein fusion protein that, when expressed from a single gene in the chromosome, restored chemotaxis to a [Delta] cheZ strain. Localization was observed in wild-type, [Delta] cheZ, [Delta] cheYZ, and [Delta] cheRB cells but not in cells with cheA, cheW, or all chemoreceptor genes except aer deleted. Cells making only CheA-short (CheA sub(S)) or CheA lacking the P2 domain also retained normal localization, whereas cells producing only CheA-long or CheA missing the P1 and P2 domains did not. We conclude that CheZ localization requires the truncated C-terminal portion of the P1 domain present in CheA sub(S). Missense mutations targeting residues 83 through 120 of CheZ also abolished localization. Two of these mutations do not disrupt chemotaxis, indicating that they specifically prevent interaction with CheA sub(S) while leaving other activities of CheZ intact. We have investigated the conditions required for polar localization of the CheZ phosphatase by using a CheZ-green fluorescent protein fusion protein that, when expressed from a single gene in the chromosome, restored chemotaxis to a cheZ strain. Localization was observed in wild-type, cheZ, cheYZ, and cheRB cells but not in cells with cheA, cheW, or all chemoreceptor genes except aer deleted. Cells making only CheA-short (CheAS) or CheA lacking the P2 domain also retained normal localization, whereas cells producing only CheA-long or CheA missing the P1 and P2 domains did not. We conclude that CheZ localization requires the truncated C-terminal portion of the P1 domain present in CheAS. Missense mutations targeting residues 83 through 120 of CheZ also abolished localization. Two of these mutations do not disrupt chemotaxis, indicating that they specifically prevent interaction with CheAS while leaving other activities of CheZ intact. [PUBLICATION ABSTRACT] We have investigated the conditions required for polar localization of the CheZ phosphatase by using a CheZ-green fluorescent protein fusion protein that, when expressed from a single gene in the chromosome, restored chemotaxis to a Δ cheZ strain. Localization was observed in wild-type, Δ cheZ , Δ cheYZ , and Δ cheRB cells but not in cells with cheA , cheW , or all chemoreceptor genes except aer deleted. Cells making only CheA-short (CheA S ) or CheA lacking the P2 domain also retained normal localization, whereas cells producing only CheA-long or CheA missing the P1 and P2 domains did not. We conclude that CheZ localization requires the truncated C-terminal portion of the P1 domain present in CheA S . Missense mutations targeting residues 83 through 120 of CheZ also abolished localization. Two of these mutations do not disrupt chemotaxis, indicating that they specifically prevent interaction with CheA S while leaving other activities of CheZ intact. Article Usage Stats Services JB Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue JB About JB Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy JB RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0021-9193 Online ISSN: 1098-5530 Copyright © 2014 by the American Society for Microbiology. For an alternate route to JB .asm.org, visit: JB |
| Author | Roger R. Draheim Michael D. Manson Brian J. Cantwell Cameran Nguyen Richard C. Stewart Richard B. Weart |
| AuthorAffiliation | Department of Biology, Texas A&M University, College Station, Texas 77843, 1 Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Maryland 20742 2 |
| AuthorAffiliation_xml | – name: Department of Biology, Texas A&M University, College Station, Texas 77843, 1 Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Maryland 20742 2 |
| Author_xml | – sequence: 1 givenname: Brian J surname: Cantwell fullname: Cantwell, Brian J organization: Department of Biology, Texas A&M University, College Station, Texas 77843, USA – sequence: 2 givenname: Roger R surname: Draheim fullname: Draheim, Roger R – sequence: 3 givenname: Richard B surname: Weart fullname: Weart, Richard B – sequence: 4 givenname: Cameran surname: Nguyen fullname: Nguyen, Cameran – sequence: 5 givenname: Richard C surname: Stewart fullname: Stewart, Richard C – sequence: 6 givenname: Michael D surname: Manson fullname: Manson, Michael D |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/12644507$$D View this record in MEDLINE/PubMed |
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Mendeley... We have investigated the conditions required for polar localization of the CheZ phosphatase by using a CheZ-green fluorescent protein fusion protein that, when... |
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| SubjectTerms | Amino Acid Sequence Bacterial Proteins - chemistry Bacterial Proteins - genetics Bacterial Proteins - metabolism Bacteriology Chemoreceptor Cells - metabolism Chemotaxis - genetics Conserved Sequence Enterobacteriaceae - metabolism Green Fluorescent Proteins Hydrophobic and Hydrophilic Interactions Luminescent Proteins - genetics Luminescent Proteins - metabolism Models, Molecular Molecular Sequence Data Mutation Phosphates Protein Conformation Protein Kinases - chemistry Protein Kinases - genetics Protein Kinases - metabolism Protein Structure, Tertiary Proteins Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Sequence Homology, Amino Acid Signal Transduction Subcellular Fractions |
| Title | CheZ Phosphatase Localizes to Chemoreceptor Patches via CheA-Short |
| URI | http://jb.asm.org/content/185/7/2354.abstract https://www.ncbi.nlm.nih.gov/pubmed/12644507 https://www.proquest.com/docview/227100778 https://search.proquest.com/docview/18691618 https://search.proquest.com/docview/73097951 https://pubmed.ncbi.nlm.nih.gov/PMC151485 |
| Volume | 185 |
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