Systematic functional comparative analysis of four single-stranded DNA-binding proteins and their affection on viral RNA metabolism.
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.
National Institute for Viral Disease Control and Prevention, China Center for Disease Control and Prevention, Beijing, China.
Haiyan
H
Shi
Yonghui
Y
Zhang
Guohui
G
Zhang
Jinlei
J
Guo
Xun
X
Zhang
Haiyan
H
Song
Jianxin
J
Lv
Jimin
J
Gao
Yuepeng
Y
Wang
Litian
L
Chen
Yue
Y
Wang
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D045744:Cell Line, Tumor; D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D004706:Endodeoxyribonucleases; D004722:Endoribonucleases; D004926:Escherichia coli / Q000737:chemistry*; D016174:Hepacivirus / Q000187:drug effects* / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects / Q000821:virology; D006358:Hot Temperature; D006801:Humans; D011485:Protein Binding; D055550:Protein Stability; D012367:RNA, Viral / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D048229:Sulfolobus solfataricus / Q000737:chemistry; D019712:Thermococcus / Q000737:chemistry; D016963:Thermus thermophilus / Q000737:chemistry
D001426:Bacterial Proteins; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D012367:RNA, Viral; D011994:Recombinant Proteins; D004706:Endodeoxyribonucleases; D004722:Endoribonucleases; C021835:Serratia marcescens nuclease