Staphylococcus aureus FtsZ and PBP4 bind to the conformationally dynamic N-terminal domain of GpsB

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Abstract

In the Firmicutes phylum, GpsB is a membrane associated protein that coordinates peptidoglycan synthesis with cell growth and division. Although GpsB has been studied in several bacteria, the structure, function, and interactome of Staphylococcus aureus GpsB is largely uncharacterized. To address this knowledge gap, we solved the crystal structure of the N-terminal domain of S. aureus GpsB, which adopts an atypical, asymmetric dimer, and demonstrates major conformational flexibility that can be mapped to a hinge region formed by a three-residue insertion exclusive to Staphylococci. When this three-residue insertion is excised, its thermal stability increases, and the mutant no longer produces a previously reported lethal phenotype when overexpressed in Bacillus subtilis. In S. aureus, we show that these hinge mutants are less functional and speculate that the conformational flexibility imparted by the hinge region may serve as a dynamic switch to finetune the function of the GpsB complex and/or to promote interaction with its various partners. Furthermore, we provide the first biochemical, biophysical, and crystallographic evidence that the N-terminal domain of GpsB binds not only PBP4, but also FtsZ, through a conserved recognition motif located on their C-termini, thus coupling peptidoglycan synthesis to cell division. Taken together, the unique structure of S. aureus GpsB and its direct interaction with FtsZ/PBP4 provide deeper insight into the central role of GpsB in S. aureus cell division.

Data availability

All crystal structures have been deposited in the RCSB Protein Data Bank (PDB) with accession IDs of: Sa GpsB NTD (PDB ID 8E2B), Sa GpsB NTD + Sa PBP4 C-term (PDB ID 8E2C).

The following data sets were generated

1. Sacco M
2. Chen Y
(2022) N-terminal domain of S. aureus GpsB
PDB, 8E2B.

https://www.rcsb.org/structure/8E2B
1. Sacco M
2. Chen Y
(2022) N-terminal domain of S. aureus GpsB in complex with PBP4 fragment
PDB, 8E2C.

https://www.rcsb.org/structure/8E2C

Article and author information

Author details

Michael D Sacco

Department of Molecular Medicine, University of South Florida, Tampa, United States

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The authors declare that no competing interests exist.
Lauren R Hammond

Department of Molecular Biosciences, University of South Florida, Tampa, United States

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Radwan E Noor

Global and Planetary Health, University of South Florida, Tampa, United States

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Dipanwita Bhattacharya

Department of Molecular Biosciences, University of South Florida, Tampa, United States

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The authors declare that no competing interests exist.
Lily J McKnight

Department of Molecular Biosciences, University of South Florida, Tampa, United States

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The authors declare that no competing interests exist.
Jesper J Madsen

Department of Molecular Medicine, University of South Florida, Tampa, United States

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The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0003-1411-9080
Xiujun Zhang

Department of Molecular Medicine, University of South Florida, Tampa, United States

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The authors declare that no competing interests exist.
Shane G Butler

Department of Molecular Medicine, University of South Florida, Tampa, United States

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M Trent Kemp

Department of Molecular Medicine, University of South Florida, Tampa, United States

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The authors declare that no competing interests exist.
Aiden C Jaskolka-Brown

Department of Molecular Medicine, University of South Florida, Tampa, United States

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The authors declare that no competing interests exist.
Sebastian J Khan

Department of Molecular Biosciences, University of South Florida, Tampa, United States

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The authors declare that no competing interests exist.
Ioannis Gelis

Department of Chemistry, University of South Florida, Tampa, United States

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The authors declare that no competing interests exist.
Prahathees Eswara

Department of Molecular Biosciences, University of South Florida, Tampa, United States

For correspondence
eswara@usf.edu

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0003-4430-261X
Yu Chen

Department of Molecular Medicine, University of South Florida, Tampa, United States

For correspondence
ychen1@usf.edu

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-5115-3600

Funding

National Institutes of Health (R21 AI164775)

Yu Chen

National Institutes of Health (R35 GM133617)

Prahathees Eswara

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.