Identification of SPOR domain amino acids important for septal localization, peptidoglycan binding, and a disulfide bond in the cell division protein FtsN

Tammi R Duncan; Atsushi Yahashiri; S J Ryan Arends; David L Popham; David S Weiss

doi:10.1128/JB.00911-13

Back

Identification of SPOR domain amino acids important for septal localization, peptidoglycan binding, and a disulfide bond in the cell division protein FtsN

Journal article

Open access

Peer reviewed

Identification of SPOR domain amino acids important for septal localization, peptidoglycan binding, and a disulfide bond in the cell division protein FtsN

Tammi R Duncan, Atsushi Yahashiri, S J Ryan Arends, David L Popham and David S Weiss

Journal of bacteriology, Vol.195(23), pp.5308-5315

12/2013

DOI: 10.1128/JB.00911-13

PMCID: PMC3837948

PMID: 24056104

Files and links (1)

url

https://doi.org/10.1128/JB.00911-13View

Published (Version of record) Open Access

Abstract

SPOR domains are about 75 amino acids long and probably bind septal peptidoglycan during cell division. We mutagenized 33 amino acids with surface-exposed side chains in the SPOR domain from an Escherichia coli cell division protein named FtsN. The mutant SPOR domains were fused to Tat-targeted green fluorescent protein ((TT)GFP) and tested for septal localization in live E. coli cells. Lesions at the following 5 residues reduced septal localization by a factor of 3 or more: Q251, S254, W283, R285, and I313. All of these residues map to a β-sheet in the published solution structure of FtsN(SPOR). Three of the mutant proteins (Q251E, S254E, and R285A mutants) were purified and found to be defective in binding to peptidoglycan sacculi in a cosedimentation assay. These results match closely with results from a previous study of the SPOR domain from DamX, even though these two SPOR domains share <20% amino acid identity. Taken together, these findings support the proposal that SPOR domains localize by binding to septal peptidoglycan and imply that the binding site is associated with the β-sheet. We also show that FtsN(SPOR) contains a disulfide bond between β-sheet residues C252 and C312. The disulfide bond contributes to protein stability, cell division, and peptidoglycan binding.

Peptidoglycan - metabolism

Amino Acid Sequence

Membrane Proteins - genetics

Models, Molecular

Molecular Sequence Data

Escherichia coli Proteins - metabolism

Protein Transport - physiology

Gene Expression Regulation, Bacterial - physiology

Cell Division - physiology

Escherichia coli - genetics

Escherichia coli - metabolism

Escherichia coli Proteins - genetics

Protein Binding

Protein Conformation

Membrane Proteins - metabolism

Mutation

Protein Structure, Tertiary - physiology

Details

Title: Subtitle: Identification of SPOR domain amino acids important for septal localization, peptidoglycan binding, and a disulfide bond in the cell division protein FtsN
Creators: Tammi R Duncan - Department of Microbiology, Carver College of Medicine, The University of Iowa, Iowa City, Iowa, USA
Atsushi Yahashiri
S J Ryan Arends
David L Popham
David S Weiss
Resource Type: Journal article
Publication Details: Journal of bacteriology, Vol.195(23), pp.5308-5315
Publisher: United States
DOI: 10.1128/JB.00911-13
PMID: 24056104
PMCID: PMC3837948
ISSN: 0021-9193
eISSN: 1098-5530
Grant note: GM083975 / NIGMS NIH HHS R01 GM083975 / NIGMS NIH HHS
Language: English
Date published: 12/2013
Academic Unit: Microbiology and Immunology
Record Identifier: 9984001208502771

Metrics

13 Record Views

18 Times Cited - Web of Science

See more details