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Autophosphorylation of the KaiC-like protein ArlH inhibits oligomerization and interaction with ArlI, the motor ATPase of the archaellum.
de Sousa Machado, J Nuno; Vollmar, Leonie; Schimpf, Julia; Chaudhury, Paushali; Kumariya, Rashmi; van der Does, Chris; Hugel, Thorsten; Albers, Sonja-Verena.
Afiliação
  • de Sousa Machado JN; Molecular Biology of Archaea and Signaling Research Centre BIOSS, Institute of Biology II, Faculty of Biology, University of Freiburg, Freiburg, Germany.
  • Vollmar L; Spemann Graduate School of Biology and Medicine, University of Freiburg, Freiburg, Germany.
  • Schimpf J; Spemann Graduate School of Biology and Medicine, University of Freiburg, Freiburg, Germany.
  • Chaudhury P; Institute of Physical Chemistry and Signaling Research Centers BIOSS and CIBSS, University of Freiburg, Freiburg, Germany.
  • Kumariya R; Spemann Graduate School of Biology and Medicine, University of Freiburg, Freiburg, Germany.
  • van der Does C; Institute of Physical Chemistry and Signaling Research Centers BIOSS and CIBSS, University of Freiburg, Freiburg, Germany.
  • Hugel T; Molecular Biology of Archaea and Signaling Research Centre BIOSS, Institute of Biology II, Faculty of Biology, University of Freiburg, Freiburg, Germany.
  • Albers SV; Molecular Biology of Archaea and Signaling Research Centre BIOSS, Institute of Biology II, Faculty of Biology, University of Freiburg, Freiburg, Germany.
Mol Microbiol ; 116(3): 943-956, 2021 09.
Article em En | MEDLINE | ID: mdl-34219289
ABSTRACT
Motile archaea are propelled by the archaellum, whose motor complex consists of the membrane protein ArlJ, the ATPase ArlI, and the ATP-binding protein ArlH. Despite its essential function and the existence of structural and biochemical data on ArlH, the role of ArlH in archaellum assembly and function remains elusive. ArlH is a structural homolog of KaiC, the central component of the cyanobacterial circadian clock. Since autophosphorylation and dephosphorylation of KaiC are central properties for the function of KaiC, we asked whether autophosphorylation is also a property of ArlH proteins. We observed that both ArlH from the euryarchaeon Pyrococcus furiosus (PfArlH) and from the crenarchaeon Sulfolobus acidocaldarius (SaArlH) have autophosphorylation activity. Using a combination of single-molecule fluorescence measurements and biochemical assays, we show that autophosphorylation of ArlH is closely linked to its oligomeric state when bound to hexameric ArlI. These experiments also strongly suggest that ArlH is a hexamer in its ArlI-bound state. Mutagenesis of the putative catalytic residue (Glu-57 in SaArlH) in ArlH results in a reduced autophosphorylation activity and abolished archaellation and motility in S. acidocaldarius, indicating that optimum phosphorylation activity of ArlH is essential for archaellation and motility.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Sulfolobus acidocaldarius / Adenosina Trifosfatases / Pyrococcus furiosus / Peptídeos e Proteínas de Sinalização do Ritmo Circadiano / Movimento Idioma: En Ano de publicação: 2021 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Sulfolobus acidocaldarius / Adenosina Trifosfatases / Pyrococcus furiosus / Peptídeos e Proteínas de Sinalização do Ritmo Circadiano / Movimento Idioma: En Ano de publicação: 2021 Tipo de documento: Article