VirE2, a Type IV secretion substrate, interacts with the VirD4 transfer protein at cell poles of Agrobacterium tumefaciens

VirE2, a Type IV secretion substrate, interacts with the VirD4 transfer protein at cell poles of... Summary Agrobacterium tumefaciens transfers oncogenic DNA and effector proteins to plant cells during the course of infection. Substrate translocation across the bacterial cell envelope is mediated by a type IV secretion (TFS) system composed of the VirB proteins, as well as VirD4, a member of a large family of inner membrane proteins implicated in the coupling of DNA transfer intermediates to the secretion machine. In this study, we demonstrate with novel cytological screens – a two‐hybrid (C2H) assay and bimolecular fluorescence complementation (BiFC) – and by immunoprecipitation of chemically cross‐linked protein complexes that the VirE2 effector protein interacts directly with the VirD4 coupling protein at cell poles of A. tumefaciens. Analyses of truncation derivatives showed that VirE2 interacts via its C terminus with VirD4, and, further, an NH2‐terminal membrane‐spanning domain of VirD4 is dispensable for complex formation. VirE2 interacts with VirD4 independently of the virB‐encoded transfer machine and T pilus, the putative periplasmic chaperones AcvB and VirJ, and the T‐DNA transfer intermediate. Finally, VirE2 is recruited to polar‐localized VirD4 as a complex with its stabilizing secretion chaperone VirE1, yet the effector–coupling protein interaction is not dependent on chaperone binding. Together, our findings establish for the first time that a protein substrate of a type IV secretion system is recruited to a member of the coupling protein superfamily. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Molecular Microbiology Wiley

VirE2, a Type IV secretion substrate, interacts with the VirD4 transfer protein at cell poles of Agrobacterium tumefaciens

Loading next page...
1
 
/lp/wiley/vire2-a-type-iv-secretion-substrate-interacts-with-the-vird4-transfer-S6n0aJEGBu
Publisher site
See Article on Publisher Site

Abstract

Summary Agrobacterium tumefaciens transfers oncogenic DNA and effector proteins to plant cells during the course of infection. Substrate translocation across the bacterial cell envelope is mediated by a type IV secretion (TFS) system composed of the VirB proteins, as well as VirD4, a member of a large family of inner membrane proteins implicated in the coupling of DNA transfer intermediates to the secretion machine. In this study, we demonstrate with novel cytological screens – a two‐hybrid (C2H) assay and bimolecular fluorescence complementation (BiFC) – and by immunoprecipitation of chemically cross‐linked protein complexes that the VirE2 effector protein interacts directly with the VirD4 coupling protein at cell poles of A. tumefaciens. Analyses of truncation derivatives showed that VirE2 interacts via its C terminus with VirD4, and, further, an NH2‐terminal membrane‐spanning domain of VirD4 is dispensable for complex formation. VirE2 interacts with VirD4 independently of the virB‐encoded transfer machine and T pilus, the putative periplasmic chaperones AcvB and VirJ, and the T‐DNA transfer intermediate. Finally, VirE2 is recruited to polar‐localized VirD4 as a complex with its stabilizing secretion chaperone VirE1, yet the effector–coupling protein interaction is not dependent on chaperone binding. Together, our findings establish for the first time that a protein substrate of a type IV secretion system is recruited to a member of the coupling protein superfamily.

Journal

Molecular MicrobiologyWiley

Published: Sep 1, 2003

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create folders to
organize your research

Export folders, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off