4APV

The Klebsiella pneumoniae primosomal PriB protein: identification, crystal structure, and ssDNA binding mode

Summary for 4APV

• Entry DOI: 10.2210/pdb4apv/pdb
• Descriptor: PRIMOSOMAL REPLICATION PROTEIN N (2 entities in total)
• Functional Keywords: replication, prib primosome ssdna binding
• Biological source: KLEBSIELLA PNEUMONIAE
• Total number of polymer chains: 1
• Total formula weight: 12559.39

Authors

Lo, Y.H.,Huang, Y.H.,Hsiao, C.D.,Huang, C.Y. (deposition date: 2012-04-06, release date: 2012-04-25, Last modification date: 2024-11-20)

Primary citation

Huang, Y.,Lo, Y.H.,Huang, W.,Huang, C.Y.
Crystal Structure and DNA-Binding Mode of Klebsiella Pneumoniae Primosomal Prib Protein.
Genes Cells, 17:837-, 2012

PubMed Abstract: PriB is a primosomal DNA replication protein required for the re-initiation of replication in bacteria. In this study, we investigated the gene expression of PriB in Klebsiella pneumoniae (KpPriB) and characterized the gene product through crystal structural and functional analyses. Quantitative polymerase chain reaction analysis (Q-PCR) indicated that the 104-aa priB was expressed in K. pneumoniae with a C(T) value of 22.4. The crystal structure of KpPriB (Protein Data Bank entry: 4APV) determined at a resolution of 2.1 Å was similar to that of Escherichia coli PriB (EcPriB). KpPriB formed a single complex with single-stranded DNA (ssDNA) of different lengths, suggesting a highly cooperative process. Structure-based mutational analysis revealed that substitution at K18, F42, R44, W47, K82, K84, or K89 but not R34 in KpPriB had a significant effect on both ssDNA and double-stranded DNA (dsDNA) binding. Based on these findings, the known ssDNA interaction sites of PriB were expanded to include R44 and F42, thus allowing nucleic acids to wrap around the whole PriB protein.

PubMed: 22938024
DOI: 10.1111/GTC.12001

Experimental method

X-RAY DIFFRACTION (2.095 Å)