RRC ID 44274
Author Shi H, Zhang Y, Zhang G, Guo J, Zhang X, Song H, Lv J, Gao J, Wang Y, Chen L, Wang Y.
Title Systematic functional comparative analysis of four single-stranded DNA-binding proteins and their affection on viral RNA metabolism.
Journal PLoS ONE
Abstract The accumulation of single-stranded DNA-binding (SSB) proteins is essential for organisms and has various applications. However, no study has simultaneously and systematically compared the characteristics of SSB proteins. In addition, SSB proteins may bind RNA and play an unknown biological role in RNA metabolism. Here, we expressed a novel species of SSB protein derived from Thermococcus kodakarensis KOD1 (KOD), as well as SSB proteins from Thermus thermophilus (TTH), Escherichia coli, and Sulfolobus Solfataricus P2 (SSOB), abbreviated kod, tth, bl21, and ssob, respectively. These SSB proteins could bind ssDNA and viral RNA. bl21 resisted heat treatment for more than 9 h, Ssob and kod could withstand 95°C for 10 h and retain its ssDNA- and RNA-binding ability. Four SSB proteins promoted the specificity of the DNA polymerase in PCR-based 5- and 9-kb genome fragment amplification. kod also increased the amplification of a 13-kb PCR product, and SSB protein-bound RNA resisted Benzonase digestion. The SSB proteins could also enter the host cell bound to RNA, which resulted in modulation of viral RNA metabolism, particularly ssob and bl21.
Volume 8(1)
Pages e55076
Published 2013
DOI 10.1371/journal.pone.0055076
PII PONE-D-12-25523
PMID 23365690
PMC PMC3554691
MeSH Bacterial Proteins / genetics Bacterial Proteins / metabolism* Bacterial Proteins / pharmacology Cell Line, Tumor DNA, Single-Stranded / chemistry* DNA, Single-Stranded / genetics DNA, Single-Stranded / metabolism DNA-Binding Proteins / genetics DNA-Binding Proteins / metabolism* DNA-Binding Proteins / pharmacology Endodeoxyribonucleases Endoribonucleases Escherichia coli / chemistry* Hepacivirus / drug effects* Hepacivirus / metabolism Hepatocytes / drug effects Hepatocytes / virology Hot Temperature Humans Protein Binding Protein Stability RNA, Viral / chemistry* RNA, Viral / genetics RNA, Viral / metabolism Recombinant Proteins / genetics Recombinant Proteins / metabolism Recombinant Proteins / pharmacology Sulfolobus solfataricus / chemistry Thermococcus / chemistry Thermus thermophilus / chemistry
IF 2.766
Times Cited 8
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells