Identification of C-terminal hydrophobic residues important for dimerization and all known functions of ParB of Pseudomonas aeruginosa.
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Publication year
2012Source
Microbiology (New York), 158, Pt 5, (2012), pp. 1183-95ISSN
Annotation
01 mei 2012
Publication type
Article / Letter to editor
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Organization
Laboratory of Genetic, Endocrine and Metabolic Diseases
Journal title
Microbiology (New York)
Volume
vol. 158
Issue
iss. Pt 5
Page start
p. 1183
Page end
p. 95
Subject
IGMD 6: Hormonal regulation ONCOL 5: Aetiology, screening and detection; Laboratory Medicine Radboud University Medical CenterAbstract
The ParB protein of Pseudomonas aeruginosa is important for growth, cell division, nucleoid segregation and different types of motility. To further understand its function we have demonstrated a vital role of the hydrophobic residues in the C terminus of ParB(P.a.). By in silico modelling of the C-terminal domain (amino acids 242-290) the hydrophobic residues L282, V285 and I289 (but not L286) are engaged in leucine-zipper-like structure formation, whereas the charged residues R290 and Q266 are implicated in forming a salt bridge involved in protein stabilization. Five parB mutant alleles were constructed and their functionality was defined in vivo and in vitro. In agreement with model predictions, the substitution L286A had no effect on mutant protein activities. Two ParBs with single substitutions L282A or V285A and deletions of two or seven C-terminal amino acids were impaired in both dimerization and DNA binding and were not able to silence genes adjacent to parS, suggesting that dimerization through the C terminus is a prerequisite for spreading on DNA. The defect in dimerization also correlated with loss of ability to interact with partner protein ParA. Reverse genetics demonstrated that a parB mutant producing ParB lacking the two C-terminal amino acids as well as mutants producing ParB with single substitution L282A or V285A had defects similar to those of a parB null mutant. Thus so far all the properties of ParB seem to depend on dimerization.
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- Faculty of Medical Sciences [93207]
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