Recruitment and activation of a lipid kinase by hepatitis C virus NS5A is essential for integrity of the membranous replication compartment.
Hepatitis C virus (HCV) is a major causative agent of chronic liver disease in humans. To gain insight into host factor requirements for HCV replication, we performed a siRNA screen of the human kinome and identified 13 different kinases, including phosphatidylinositol-4 kinase III alpha (PI4KIIIα), as being required for HCV replication. Consistent with elevated levels of the PI4KIIIα product phosphatidylinositol-4-phosphate (PI4P) detected in HCV-infected cultured hepatocytes and liver tissue from chronic hepatitis C patients, the enzymatic activity of PI4KIIIα was critical for HCV replication. Viral nonstructural protein 5A (NS5A) was found to interact with PI4KIIIα and stimulate its kinase activity. The absence of PI4KIIIα activity induced a dramatic change in the ultrastructural morphology of the membranous HCV replication complex. Our analysis suggests that the direct activation of a lipid kinase by HCV NS5A contributes critically to the integrity of the membranous viral replication complex.
The Department of Infectious Diseases, Molecular Virology, Heidelberg University, 69120 Heidelberg, Germany.
Simon
S
Reiss
Ilka
I
Rebhan
Perdita
P
Backes
Ines
I
Romero-Brey
Holger
H
Erfle
Petr
P
Matula
Lars
L
Kaderali
Marion
M
Poenisch
Hagen
H
Blankenburg
Marie-Sophie
MS
Hiet
Thomas
T
Longerich
Sarah
S
Diehl
Fidel
F
Ramirez
Tamas
T
Balla
Karl
K
Rohr
Artur
A
Kaul
Sandra
S
Bühler
Rainer
R
Pepperkok
Thomas
T
Lengauer
Mario
M
Albrecht
Roland
R
Eils
Peter
P
Schirmacher
Volker
V
Lohmann
Ralf
R
Bartenschlager
D002460:Cell Line; D055785:Gene Knockdown Techniques; D016174:Hepacivirus / Q000502:physiology*; D022781:Hepatocytes / Q000737:chemistry / Q000201:enzymology / Q000821:virology; D054884:Host-Pathogen Interactions; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000201:enzymology / Q000821:virology; D015778:Minor Histocompatibility Antigens; D008954:Models, Biological; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism*; D011485:Protein Binding; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D014779:Virus Replication; D012324:RNA-Dependent RNA Polymerase
D015778:Minor Histocompatibility Antigens; D018129:Phosphatidylinositol Phosphates; D017853:Phosphotransferases (Alcohol Group Acceptor); D034741:RNA, Small Interfering; D017361:Viral Nonstructural Proteins; C037178:phosphatidylinositol 4-phosphate; C402182:phosphatidylinositol phosphate 4-kinase; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase
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