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C-Terminal Domain Swapping of SSB Changes the Size of the ssDNA Binding Site
Joint Authors
Huang, Cheng-Yang
Huang, Yen-Hua
Source
Issue
Vol. 2014, Issue 2014 (31 Dec. 2014), pp.1-16, 16 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2014-08-04
Country of Publication
Egypt
No. of Pages
16
Main Subjects
Abstract EN
Single-stranded DNA-binding protein (SSB) plays an important role in DNA metabolism, including DNA replication, repair, and recombination, and is therefore essential for cell survival.
Bacterial SSB consists of an N-terminal ssDNA-binding/oligomerization domain and a flexible C-terminal protein-protein interaction domain.
We characterized the ssDNA-binding properties of Klebsiella pneumoniae SSB (KpSSB), Salmonella enterica Serovar Typhimurium LT2 SSB (StSSB), Pseudomonas aeruginosa PAO1 SSB (PaSSB), and two chimeric KpSSB proteins, namely, KpSSBnStSSBc and KpSSBnPaSSBc.
The C-terminal domain of StSSB or PaSSB was exchanged with that of KpSSB through protein chimeragenesis.
By using the electrophoretic mobility shift assay, we characterized the stoichiometry of KpSSB, StSSB, PaSSB, KpSSBnStSSBc, and KpSSBnPaSSBc, complexed with a series of ssDNA homopolymers.
The binding site sizes were determined to be 26±2, 21±2, 29±2, 21±2, and 29±2 nucleotides (nt), respectively.
Comparison of the binding site sizes of KpSSB, KpSSBnStSSBc, and KpSSBnPaSSBc showed that the C-terminal domain swapping of SSB changes the size of the binding site.
Our observations suggest that not only the conserved N-terminal domain but also the C-terminal domain of SSB is an important determinant for ssDNA binding.
American Psychological Association (APA)
Huang, Yen-Hua& Huang, Cheng-Yang. 2014. C-Terminal Domain Swapping of SSB Changes the Size of the ssDNA Binding Site. BioMed Research International،Vol. 2014, no. 2014, pp.1-16.
https://search.emarefa.net/detail/BIM-481912
Modern Language Association (MLA)
Huang, Yen-Hua& Huang, Cheng-Yang. C-Terminal Domain Swapping of SSB Changes the Size of the ssDNA Binding Site. BioMed Research International No. 2014 (2014), pp.1-16.
https://search.emarefa.net/detail/BIM-481912
American Medical Association (AMA)
Huang, Yen-Hua& Huang, Cheng-Yang. C-Terminal Domain Swapping of SSB Changes the Size of the ssDNA Binding Site. BioMed Research International. 2014. Vol. 2014, no. 2014, pp.1-16.
https://search.emarefa.net/detail/BIM-481912
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-481912