Effect of NS3 and NS5B proteins on classical swine fever virus internal ribosome entry site-mediated translation and its host cellular translation.
A full-length NS3 (NS3F) and a truncated NS3 protein (NS3H) with an RNA helicase domain possess RNA helicase activity. Using an in vitro system with a monocistronic reporter RNA or DNA, containing the CSFV 5'-UTR, we observed that both NS3F and NS3H enhanced internal ribosome entry site (IRES)-mediated and cellular translation in a dose-dependent manner, but NS3 protease (NS3P) that lacks a helicase domain did not. NS3F was stronger than NS3H in promoting both translations. These results showed that viral RNA helicase could promote viral and cellular translation, and higher RNA helicase activity might be more efficient. The NS5B protein, the viral replicase, did not significantly affect the IRES-directed or cellular translation alone. NS5B significantly enhanced the stimulative effect of NS3F on both IRES-mediated and cellular translation, but did not affect that of NS3H or NS3P. This suggests that NS5B and NS3 interact via the protease domain during the enhancement of translation.
Xiao
Ming
M
College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, PR China.
Bai
Yan
Y
College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, PR China.
Xu
Hui
H
College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, PR China.
Geng
Xiaolu
X
College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, PR China.
Chen
Jun
J
College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, PR China.
Wang
Yujing
Y
College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, PR China.
Chen
Jiakuan
J
Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, The Institute of Biodiversity Science, Fudan University, Shanghai 200433, PR China.
Li
Bo
B
Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, The Institute of Biodiversity Science, Fudan University, Shanghai 200433, PR China.
D020121:5' Untranslated Regions / Q000737:chemistry / Q000235:genetics; D000818:Animals; D002460:Cell Line; D006691:Classical Swine Fever / Q000821:virology*; D006692:Classical Swine Fever Virus / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D020365:RNA Helicases / Q000737:chemistry / Q000502:physiology*; D012324:RNA-Dependent RNA Polymerase / Q000502:physiology; D012270:Ribosomes / Q000821:virology*; D012697:Serine Endopeptidases / Q000737:chemistry / Q000502:physiology; D013552:Swine; D015854:Up-Regulation; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000502:physiology*; D014779:Virus Replication; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D020121:5' Untranslated Regions; D020365:RNA Helicases; D012324:RNA-Dependent RNA Polymerase; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C096041:NS5 protein, flavivirus