An α-Helical Core Encodes the Dual Functions of the Chlamydial Protein IncA

Ronzone, Erik; Wesolowski, Jordan; Bauler, Laura; Bhardwaj, Anshul; Hackstadt, Ted; Paumet, Fabienne

doi:10.1074/jbc.m114.592063

Cited by 22 publications

(25 citation statements)

References 49 publications

(67 reference statements)

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“…In addition to promoting homotypic fusion of C. trachomatis inclusions, wild-type IncA and IncA core also inhibit SNAREmediated membrane fusion (23). The C. trachomatis L2 incA::bla mutant grew at a normal rate regardless of the presence or absence of IncA constructs that did or did not restore fusogenicity of the inclusion.…”

Section: Discussionmentioning

confidence: 97%

“…In contrast, overexpression of either SLD-1 or SLD-2 acted in a dominant-negative fashion to block homotypic fusion with a concomitant increase in the percentage of cells exhibiting multiple, nonfused inclusions (23). Here, we used a group II intron mutant of IncA to evaluate IncA function in the absence of a background of wild-type IncA.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, a functional domain, encompassing SLD-1 and part of SLD-2, has been identified (23,24). However, it has not been determined whether this domain is necessary and sufficient for promoting inclusion fusion in C. trachomatis without IncA in a wild-type background (23,24).…”

mentioning

confidence: 99%

“…IncA is exposed on the cytosolic face of the inclusion membrane (6) and promotes homotypic fusion of inclusions but also inhibits SNARE-mediated membrane fusion, possibly to avoid fusion with endocytic compartments (6,(22)(23)(24). Recently, a functional domain, encompassing SLD-1 and part of SLD-2, has been identified (23,24). However, it has not been determined whether this domain is necessary and sufficient for promoting inclusion fusion in C. trachomatis without IncA in a wild-type background (23,24).…”

mentioning

confidence: 99%

“…IncA, an ϳ30-kDa inclusion membrane protein, possesses two putative ␣-helical SNARE-like domains, termed SNARE-like domain-1 (SLD-1) and SNARE-like domain 2 (SLD-2), that functionally mimic eukaryotic SNARE motifs (22)(23)(24). IncA is exposed on the cytosolic face of the inclusion membrane (6) and promotes homotypic fusion of inclusions but also inhibits SNARE-mediated membrane fusion, possibly to avoid fusion with endocytic compartments (6,(22)(23)(24). Recently, a functional domain, encompassing SLD-1 and part of SLD-2, has been identified (23,24).…”

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confidence: 99%

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A Functional Core of IncA Is Required for Chlamydia trachomatis Inclusion Fusion

et al. 2016

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Chlamydia trachomatis is an obligate intracellular pathogen that is the etiological agent of a variety of human diseases, including blinding trachoma and sexually transmitted infections. Chlamydiae replicate within a membrane-bound compartment, termed an inclusion, which they extensively modify by the insertion of type III secreted proteins called Inc proteins. IncA is an inclusion membrane protein that encodes two coiled-coil domains that are homologous to eukaryotic SNARE (soluble N-ethylmaleimide-sensitive factor attachment receptor) motifs. Recent biochemical evidence suggests that a functional core, composed of SNARE-like domain 1 (SLD-1) and part of SNARE-like domain 2 (SLD-2), is required for the characteristic homotypic fusion of C. trachomatis inclusions in multiply infected cells. To verify the importance of IncA in homotypic fusion in Chlamydia, we generated an incA::bla mutant. Insertional inactivation of incA resulted in the formation of nonfusogenic inclusions, a phenotype that was completely rescued by complementation with full-length IncA. Rescue of homotypic inclusion fusion was dependent on the presence of the functional core consisting of SLD-1 and part of SLD-2. Collectively, these results confirm in vitro membrane fusion assays identifying functional domains of IncA and expand the genetic tools available for identification of chlamydia with a method for complementation of site-specific mutants.

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Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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A Functional Core of IncA Is Required for Chlamydia trachomatis Inclusion Fusion

et al. 2016

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Both the N- and C- terminal regions of the Chlamydial inclusion protein D (IncD) are required for interaction with the pleckstrin homology domain of the ceramide transport protein CERT

Kumagai

Elwell

Ando

et al. 2018

Biochemical and Biophysical Research Communications

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An obligate intracellular bacterium Chlamydia trachomatis requires host lipid ceramide for their replication within an intracellular membranous compartment, the inclusion. Chlamydial inclusion membrane protein D (IncD) composed of two closely linked long hydrophobic domains with their N- and C-termini exposed to the host cytosol, binds directly to the pleckstrin homology (PH) domain of the ceramide transport protein (CERT), likely redirecting ceramide to the inclusion. The precise regions of IncD required for this interaction have not been delineated. Using co-transfection studies together with phylogenetic studies, we demonstrate that both the IncD N- and C-terminal regions are required for binding to the CERT PH domain and define key interaction residues. Native gel electrophoresis analysis demonstrates that the transmembrane region of IncD forms SDS-resistant but dithiothreitol-sensitive homodimers, which in turn can assemble to form higher order oligomers through additional N- and C-terminal domain contacts. We propose that IncD oligomerization may facilitate high affinity binding to CERT, allowing C. trachomatis to efficiently redirect host ceramide to the inclusion.

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Deep comparative genomics among Chlamydia trachomatis lymphogranuloma venereum isolates highlights genes potentially involved in pathoadaptation

Borges

Gomes

2015

Infection, Genetics and Evolution

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An α-Helical Core Encodes the Dual Functions of the Chlamydial Protein IncA

Cited by 22 publications

References 49 publications

A Functional Core of IncA Is Required for Chlamydia trachomatis Inclusion Fusion

A Functional Core of IncA Is Required for Chlamydia trachomatis Inclusion Fusion

Both the N- and C- terminal regions of the Chlamydial inclusion protein D (IncD) are required for interaction with the pleckstrin homology domain of the ceramide transport protein CERT

Deep comparative genomics among Chlamydia trachomatis lymphogranuloma venereum isolates highlights genes potentially involved in pathoadaptation

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