DOI: 10.1128/jb.186.6.1658-1666.2004. The <i>Legionella pneumophila</i> PilT Homologue DotB Exhibits ATPase Activity That Is Critical for Intracellular Growth

The Legionella pneumophila PilT Homologue DotB Exhibits ATPase Activity That Is Critical for Intracellular Growth

10.1128/jb.186.6.1658-1666.2004

Crossref journal-article

American Society for Microbiology

Journal of Bacteriology (235)

Abstract

ABSTRACT The ability of Legionella pneumophila to grow and cause disease in the host is completely dependent on a type IV secretion system known as the Dot/Icm complex. This membrane-spanning apparatus translocates effector molecules into host cells in a process that is poorly understood but that is known to require the putative ATPase DotB. One possible role for DotB is suggested by its similarity to the PilT family of proteins, which mediate pilus retraction. To better understand the molecular behavior of DotB, we have purified the protein and shown that it forms stable homohexameric rings and hydrolyzes ATP with a specific activity of 6.4 nmol of ATP/min/mg of protein. ATPase activity is critical to the function of DotB, as alteration of the conserved Walker box lysine residue resulted in a mutant protein, DotB K162Q, which failed to bind or hydrolyze ATP and which could not complement a Δ dotB strain for intracellular growth in macrophages. Consistent with the ability of DotB to interact with itself, the dotBK162Q allele exhibited transdominance over wild-type dotB , providing the first example of such a mutation in L. pneumophila . Finally, the DotB K162Q mutant protein had a significantly enhanced membrane localization in L. pneumophila compared to wild-type DotB, suggesting a relationship between nucleotide binding and membrane association. These results are consistent with a model in which DotB cycles between the cytoplasm and the Dot/Icm complex at the membrane, where it hydrolyzes nucleotides to provide energy to the complex.

Bibliography

Sexton, J. A., Pinkner, J. S., Roth, R., Heuser, J. E., Hultgren, S. J., & Vogel, J. P. (2004). The Legionella pneumophila PilT Homologue DotB Exhibits ATPase Activity That Is Critical for Intracellular Growth. Journal of Bacteriology, 186(6), 1658â1666.

Authors 6

Jessica A. Sexton (first)
Jerome S. Pinkner (additional)
Robyn Roth (additional)
John E. Heuser (additional)
Scott J. Hultgren (additional)
Joseph P. Vogel (additional)

References 42 Referenced 127

Dates

Type	When
Created	21 years, 6 months ago (March 2, 2004, 1:12 p.m.)
Deposited	4 years, 1 month ago (July 29, 2021, 1:20 p.m.)
Indexed	4 months, 3 weeks ago (April 15, 2025, 7:31 a.m.)
Issued	21 years, 5 months ago (March 15, 2004)
Published	21 years, 5 months ago (March 15, 2004)
Published Print	21 years, 5 months ago (March 15, 2004)

Funders 0

None

BibTeX

@article{Sexton_2004, title={The Legionella pneumophila PilT Homologue DotB Exhibits ATPase Activity That Is Critical for Intracellular Growth}, volume={186}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.186.6.1658-1666.2004}, DOI={10.1128/jb.186.6.1658-1666.2004}, number={6}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Sexton, Jessica A. and Pinkner, Jerome S. and Roth, Robyn and Heuser, John E. and Hultgren, Scott J. and Vogel, Joseph P.}, year={2004}, month=mar, pages={1658–1666} }

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