Conserved residues are critical for Haloferax volcanii archaeosortase catalytic activity: Implications for convergent evolution of the catalytic mechanisms of non-homologous sortases from archaea and bacteria

Abdul Halim, MF; Rodriguez, R; Stoltzfus, JD; Duggin, IG; Pohlschroder, M

Conserved residues are critical for Haloferax volcanii archaeosortase catalytic activity: Implications for convergent evolution of the catalytic mechanisms of non-homologous sortases from archaea and bacteria

Abdul Halim, MF Rodriguez, R Stoltzfus, JD Duggin, IG

Pohlschroder, M

Permalink

Publication Type:: Journal Article
Citation:: Molecular Microbiology, 2018, 108 (3), pp. 276 - 287
Issue Date:: 2018-05-01

Closed Access

	Filename	Description	Size
	Halim_et_al-2018-Molecular_Microbiology.pdf	Accepted Manuscript Version	839.13 kB	Adobe PDF	View/Open

Copyright Clearance Process

Recently Added
In Progress
Closed Access

This item is closed access and not available.

Full metadata record

Field	Value	Language
dc.contributor.author	Abdul Halim, MF	en_US
dc.contributor.author	Rodriguez, R	en_US
dc.contributor.author	Stoltzfus, JD	en_US
dc.contributor.author	Duggin, IG https://orcid.org/0000-0003-4868-7350	en_US
dc.contributor.author	Pohlschroder, M	en_US
dc.date.available	2018-02-17	en_US
dc.date.issued	2018-05-01	en_US
dc.identifier.citation	Molecular Microbiology, 2018, 108 (3), pp. 276 - 287	en_US
dc.identifier.issn	0950-382X	en_US
dc.identifier.uri	http://hdl.handle.net/10453/124071
dc.description.abstract	© 2018 John Wiley & Sons Ltd Proper protein anchoring is key to the biogenesis of prokaryotic cell surfaces, dynamic, resilient structures that play crucial roles in various cell processes. A novel surface protein anchoring mechanism in Haloferax volcanii depends upon the peptidase archaeosortase A (ArtA) processing C-termini of substrates containing C-terminal tripartite structures and anchoring mature substrates to the cell membrane via intercalation of lipid-modified C-terminal amino acid residues. While this membrane protein lacks clear homology to soluble sortase transpeptidases of Gram-positive bacteria, which also process C-termini of substrates whose C-terminal tripartite structures resemble those of ArtA substrates, archaeosortases do contain conserved cysteine, arginine and arginine/histidine/asparagine residues, reminiscent of His-Cys-Arg residues of sortase catalytic sites. The study presented here shows that ArtAWT-GFP expressed in trans complements ΔartA growth and motility phenotypes, while alanine substitution mutants, Cys173 (C173A), Arg214 (R214A) or Arg253 (R253A), and the serine substitution mutant for Cys173 (C173S), fail to complement these phenotypes. Consistent with sortase active site replacement mutants, ArtAC173A-GFP, ArtAC173S-GFP and ArtAR214A-GFP cannot process substrates, while replacement of the third residue, ArtAR253A-GFP retains some processing activity. These findings support the view that similarities between certain aspects of the structures and functions of the sortases and archaeosortases are the result of convergent evolution.	en_US
dc.relation	http://purl.org/au-research/grants/arc/FT160100010
dc.relation.ispartof	Molecular Microbiology	en_US
dc.relation.isbasedon	10.1111/mmi.13935	en_US
dc.subject.classification	Microbiology	en_US
dc.subject.mesh	Bacteria	en_US
dc.subject.mesh	Archaea	en_US
dc.subject.mesh	Haloferax volcanii	en_US
dc.subject.mesh	Cysteine	en_US
dc.subject.mesh	Cysteine Endopeptidases	en_US
dc.subject.mesh	Aminoacyltransferases	en_US
dc.subject.mesh	Arginine	en_US
dc.subject.mesh	Histidine	en_US
dc.subject.mesh	Archaeal Proteins	en_US
dc.subject.mesh	Bacterial Proteins	en_US
dc.subject.mesh	Evolution, Molecular	en_US
dc.subject.mesh	Protein Processing, Post-Translational	en_US
dc.subject.mesh	Amino Acid Sequence	en_US
dc.subject.mesh	Catalytic Domain	en_US
dc.subject.mesh	Conserved Sequence	en_US
dc.subject.mesh	Catalysis	en_US
dc.subject.mesh	Biological Evolution	en_US
dc.title	Conserved residues are critical for Haloferax volcanii archaeosortase catalytic activity: Implications for convergent evolution of the catalytic mechanisms of non-homologous sortases from archaea and bacteria	en_US
dc.type	Journal Article
utslib.citation.volume	3	en_US
utslib.citation.volume	108	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	1101 Medical Biochemistry and Metabolomics	en_US
utslib.for	0707 Veterinary Sciences	en_US
utslib.for	06 Biological Sciences	en_US
utslib.for	07 Agricultural and Veterinary Sciences	en_US
utslib.for	11 Medical and Health Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	closed_access
pubs.issue	3	en_US
pubs.publication-status	Published	en_US
pubs.volume	108	en_US

Abstract:

© 2018 John Wiley & Sons Ltd Proper protein anchoring is key to the biogenesis of prokaryotic cell surfaces, dynamic, resilient structures that play crucial roles in various cell processes. A novel surface protein anchoring mechanism in Haloferax volcanii depends upon the peptidase archaeosortase A (ArtA) processing C-termini of substrates containing C-terminal tripartite structures and anchoring mature substrates to the cell membrane via intercalation of lipid-modified C-terminal amino acid residues. While this membrane protein lacks clear homology to soluble sortase transpeptidases of Gram-positive bacteria, which also process C-termini of substrates whose C-terminal tripartite structures resemble those of ArtA substrates, archaeosortases do contain conserved cysteine, arginine and arginine/histidine/asparagine residues, reminiscent of His-Cys-Arg residues of sortase catalytic sites. The study presented here shows that ArtAWT-GFP expressed in trans complements ΔartA growth and motility phenotypes, while alanine substitution mutants, Cys173 (C173A), Arg214 (R214A) or Arg253 (R253A), and the serine substitution mutant for Cys173 (C173S), fail to complement these phenotypes. Consistent with sortase active site replacement mutants, ArtAC173A-GFP, ArtAC173S-GFP and ArtAR214A-GFP cannot process substrates, while replacement of the third residue, ArtAR253A-GFP retains some processing activity. These findings support the view that similarities between certain aspects of the structures and functions of the sortases and archaeosortases are the result of convergent evolution.

Please use this identifier to cite or link to this item:

http://hdl.handle.net/10453/124071