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Title: The nucleotide-dependent interaction of FlaH and FlaI is essential for assembly and function of the archaellum motor

Abstract

The motor of the membrane-anchored archaeal motility structure, the archaellum, contains FlaX, FlaI and FlaH. FlaX forms a 30 nm ring structure that acts as a scaffold protein and was shown to interact with the bifunctional ATPase FlaI and FlaH. However, the structure and function of FlaH has been enigmatic. Here we present structural and functional analyses of isolated FlaH and archaellum motor subcomplexes. The FlaH crystal structure reveals a RecA/Rad51 family fold with an ATP bound on a conserved and exposed surface, which presumably forms an oligomerization interface. FlaH does not hydrolyze ATP in vitro, but ATP binding to FlaH is essential for its interaction with FlaI and for archaellum assembly. FlaH interacts with the C-terminus of FlaX, which was earlier shown to be essential for FlaX ring formation and to mediate interaction with FlaI. Electron microscopy reveals that FlaH assembles as a second ring inside the FlaX ring in vitro. Collectively these data reveal central structural mechanisms for FlaH interactions in mediating archaellar assembly: FlaH binding within the FlaX ring and nucleotide-regulated FlaH binding to FlaI form the archaellar basal body core.

Authors:
 [1];  [1];  [2];  [3];  [4];  [1];  [4];  [2];  [1];  [5];  [2];  [1]
  1. Univ. of Freiburg (Germany)
  2. Max Planck Inst. of Biophysics, Frankfurt (Germany)
  3. Philipps Univ. of Marburg (Germany)
  4. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  5. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of Texas M. D. Anderson Cancer Center, Houston, TX (United States)
Publication Date:
Research Org.:
Univ. of Freiburg (Germany); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE; National Inst. of Health (NIH) (United States); Max Planck Society (Germany); European Research Council (ERC)
OSTI Identifier:
1378738
Grant/Contract Number:  
AC02-05CH11231; GM105404; 311523
Resource Type:
Accepted Manuscript
Journal Name:
Molecular microbiology
Additional Journal Information:
Journal Volume: 99; Journal Issue: 4; Journal ID: ISSN 0950-382X
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Chaudhury, Paushali, Neiner, Tomasz, D'Imprima, Edoardo, Banerjee, Ankan, Reindl, Sophia, Ghosh, Abhrajyoti, Arvai, Andrew S., Mills, Deryck J., van der Does, Chris, Tainer, John A., Vonck, Janet, and Albers, Sonja-Verena. The nucleotide-dependent interaction of FlaH and FlaI is essential for assembly and function of the archaellum motor. United States: N. p., 2015. Web. doi:10.1111/mmi.13260.
Chaudhury, Paushali, Neiner, Tomasz, D'Imprima, Edoardo, Banerjee, Ankan, Reindl, Sophia, Ghosh, Abhrajyoti, Arvai, Andrew S., Mills, Deryck J., van der Does, Chris, Tainer, John A., Vonck, Janet, & Albers, Sonja-Verena. The nucleotide-dependent interaction of FlaH and FlaI is essential for assembly and function of the archaellum motor. United States. doi:10.1111/mmi.13260.
Chaudhury, Paushali, Neiner, Tomasz, D'Imprima, Edoardo, Banerjee, Ankan, Reindl, Sophia, Ghosh, Abhrajyoti, Arvai, Andrew S., Mills, Deryck J., van der Does, Chris, Tainer, John A., Vonck, Janet, and Albers, Sonja-Verena. Wed . "The nucleotide-dependent interaction of FlaH and FlaI is essential for assembly and function of the archaellum motor". United States. doi:10.1111/mmi.13260. https://www.osti.gov/servlets/purl/1378738.
@article{osti_1378738,
title = {The nucleotide-dependent interaction of FlaH and FlaI is essential for assembly and function of the archaellum motor},
author = {Chaudhury, Paushali and Neiner, Tomasz and D'Imprima, Edoardo and Banerjee, Ankan and Reindl, Sophia and Ghosh, Abhrajyoti and Arvai, Andrew S. and Mills, Deryck J. and van der Does, Chris and Tainer, John A. and Vonck, Janet and Albers, Sonja-Verena},
abstractNote = {The motor of the membrane-anchored archaeal motility structure, the archaellum, contains FlaX, FlaI and FlaH. FlaX forms a 30 nm ring structure that acts as a scaffold protein and was shown to interact with the bifunctional ATPase FlaI and FlaH. However, the structure and function of FlaH has been enigmatic. Here we present structural and functional analyses of isolated FlaH and archaellum motor subcomplexes. The FlaH crystal structure reveals a RecA/Rad51 family fold with an ATP bound on a conserved and exposed surface, which presumably forms an oligomerization interface. FlaH does not hydrolyze ATP in vitro, but ATP binding to FlaH is essential for its interaction with FlaI and for archaellum assembly. FlaH interacts with the C-terminus of FlaX, which was earlier shown to be essential for FlaX ring formation and to mediate interaction with FlaI. Electron microscopy reveals that FlaH assembles as a second ring inside the FlaX ring in vitro. Collectively these data reveal central structural mechanisms for FlaH interactions in mediating archaellar assembly: FlaH binding within the FlaX ring and nucleotide-regulated FlaH binding to FlaI form the archaellar basal body core.},
doi = {10.1111/mmi.13260},
journal = {Molecular microbiology},
number = 4,
volume = 99,
place = {United States},
year = {2015},
month = {10}
}

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