skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Structural and Functional Analysis of BipA, a Regulator of Virulence in Enteropathogenic Escherichia coli

Journal Article · · Journal of Biological Chemistry
 [1];  [1];  [1];  [1];  [1]
  1. Univ. of California, Riverside, CA (United States). Dept. of Biochemistry
+ Show Author Affiliations

The translational GTPase BipA regulates the expression of virulence and pathogenicity factors in several eubacteria. BipA-dependent expression of virulence factors occurs under starvation conditions, such as encountered during infection of a host. Under these conditions, BipA associates with the small ribosomal subunit. BipA also has a second function to promote the efficiency of late steps in biogenesis of large ribosomal subunits at low temperatures, presumably while bound to the ribosome. During starvation, the cellular concentration of stress alarmone guanosine-3', 5'-bis pyrophosphate (ppGpp) is increased. This increase allows ppGpp to bind to BipA and switch its binding specificity from ribosomes to small ribosomal subunits. A conformational change of BipA upon ppGpp binding could explain the ppGpp regulation of the binding specificity of BipA. Here, we present the structures of the full-length BipA from Escherichia coli in apo, GDP-, and ppGpp-bound forms. The crystal structure and small-angle x-ray scattering data of the protein with bound nucleotides, together with a thermodynamic analysis of the binding of GDP and of ppGpp to BipA, indicate that the ppGpp-bound form of BipA adopts the structure of the GDP form. This suggests furthermore, that the switch in binding preference only occurs when both ppGpp and the small ribosomal subunit are present. Finally, this molecular mechanism would allow BipA to interact with both the ribosome and the small ribosomal subunit during stress response.

Research Organization:
Univ. of California, Riverside, CA (United States)
Sponsoring Organization:
USDOE; National Inst. of Health (NIH) (United States); Univ. of California, Riverside, CA (United States)
Grant/Contract Number:
AC02-06CH11357; P41 GM103403
OSTI ID:
1212947
Journal Information:
Journal of Biological Chemistry, Vol. 290, Issue 34; ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular BiologyCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 24 works
Citation information provided by
Web of Science

References (38)

Thermodynamics of GTP and GDP Binding to Bacterial Initiation Factor 2 Suggests Two Types of Structural Transitions journal December 2009
Thermodynamic Characterization of ppGpp Binding to EF-G or IF2 and of Initiator tRNA Binding to Free IF2 in the Presence of GDP, GTP, or ppGpp journal October 2010
Regulation of the Escherichia coli K5 Capsule Gene Cluster: Evidence for the Roles of H-NS, BipA, and Integration Host Factor in Regulation of Group 2 Capsule Gene Clusters in Pathogenic E. coli journal May 2000
Implementation and performance of SIBYLS: a dual endstation small-angle X-ray scattering and macromolecular crystallography beamline at the Advanced Light Source journal January 2013
A Novel Domain in Translational GTPase BipA Mediates Interaction with the 70S Ribosome and Influences GTP Hydrolysis journal November 2009
The pretranslocation ribosome is targeted by GTP-bound EF-G in partially activated form journal October 2008
The nucleotide-binding site of bacterial translation initiation factor 2 (IF2) as a metabolic sensor journal September 2006
A dedicated translation factor controls the synthesis of the global regulator Fis journal August 2004
ppGpp is the major source of growth rate control in E. coli journal October 2010
Salmonella enterica Serovar Typhimurium BipA Exhibits Two Distinct Ribosome Binding Modes journal July 2008
Phylogenetic distribution of translational GTPases in bacteria journal January 2007
Features and development of Coot journal March 2010
Protein production by auto-induction in high-density shaking cultures journal May 2005
Efficient Assembly of Ribosomes Is Inhibited by Deletion of bipA in Escherichia coli journal March 2015
REFMAC 5 for the refinement of macromolecular crystal structures journal March 2011
Crystallographic studies of elongation factor G journal December 1995
Cofactor Dependent Conformational Switching of GTPases journal August 2008
PHENIX: a comprehensive Python-based system for macromolecular structure solution journal January 2010
Co-ordination of pathogenicity island expression by the BipA GTPase in enteropathogenic Escherichia coli (EPEC): BipA control of EPEC and EHEC virulence genes journal April 2003
The structure of LepA, the ribosomal back translocase journal March 2008
Suppression of ΔbipA Phenotypes in Escherichia coli by Abolishment of Pseudouridylation at Specific Sites on the 23S rRNA journal September 2008
Three-dimensional structure of the ribosomal translocase: elongation factor G from Thermus thermophilus. journal August 1994
Crystal structure of a mutant elongation factor G trapped with a GTP analogue journal July 2005
Crystal structure of elongation factor 4 bound to a clockwise ratcheted ribosome journal August 2014
NMRPipe: A multidimensional spectral processing system based on UNIX pipes journal November 1995
Classification and evolution of P-loop GTPases and related ATPases journal March 2002
BipA: a tyrosine-phosphorylated GTPase that mediates interactions between enteropathogenic Escherichia coli (EPEC) and epithelial cells journal April 1998
Developing advanced X-ray scattering methods combined with crystallography and computation journal March 2013
ppGpp with DksA controls gene expression in the locus of enterocyte effacement (LEE) pathogenicity island of enterohaemorrhagic Escherichia coli through activation of two virulence regulatory genes: Stringent response system in EHEC pathogenicity journal May 2006
MOLREP an Automated Program for Molecular Replacement journal December 1997
The GTPase superfamily: conserved structure and molecular mechanism journal January 1991
The structure of elongation factor G in complex with GDP: conformational flexibility and nucleotide exchange journal May 1996
Structure of a mutant EF-G reveals domain III and possibly the fusidic acid binding site 1 1Edited by I. A. Wilson journal November 2000
The Conformational Properties of Elongation Factor G and the Mechanism of Translocation † journal August 1997
ppGpp Conjures Bacterial Virulence journal June 2010
Phaser crystallographic software journal July 2007
Conformational Changes of Elongation Factor G on the Ribosome during tRNA Translocation journal January 2015
[20] Processing of X-ray diffraction data collected in oscillation mode book January 1997

Cited By (3)

Evolution of (p)ppGpp-HPRT regulation through diversification of an allosteric oligomeric interaction journal September 2019
Genome-wide effects on Escherichia coli transcription from ppGpp binding to its two sites on RNA polymerase journal April 2019
The magic dance of the alarmones (p)ppGpp: The structural biology of the alarmones (p)ppGpp journal June 2016

Similar Records

Salmonella Enterica Serovar Typhimurium BipA Exhibits Two Distinct Ribosome Binding Modes
Journal Article · Tue Jan 01 00:00:00 EST 2008 · Journal of Bacteriology · OSTI ID:1212947
An Allosteric Pathway Revealed in the Ribosome Binding Stress Factor BipA
Journal Article · Thu Jan 01 00:00:00 EST 2009 · Biophysical Journal · OSTI ID:1212947
Dissection of the molecular circuitry controlling virulence in Francisella tularensis
Journal Article · Fri Sep 01 00:00:00 EDT 2017 · Genes & Development · OSTI ID:1212947
+4 more

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
small GTPase
structural biology
structure-function
thermodynamics
translation regulation
translational regulation