PMID- 10488152
OWN - NLM
STAT- MEDLINE
DCOM- 19991104
LR  - 20220110
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 39
DP  - 1999 Sep 24
TI  - Identification of the major phosphorylation site of the hepatitis C virus H
      strain NS5A protein as serine 2321.
PG  - 28011-8
AB  - The hepatitis C virus (HCV) NS5A protein is phosphorylated by a cellular,
      serine/threonine kinase. To identify the major site(s) of NS5A phosphorylation,
      radiolabeled HCV-H NS5A phosphopeptides were purified and subjected to
      phosphoamino acid analysis and Edman degradation. These data identified the major
      intracellular phosphorylation site in the HCV-H NS5A protein as Ser(2321), a
      result verified by two additional, independent methods: (i) substitution of Ala
      for Ser(2321) and the concomitant disappearance of the major in vivo
      phosphorylated peptides and corresponding in vitro phosphorylated peptides; and
      (ii) comigration of the digestion products of a synthetic peptide phosphorylated 
      on Ser(2321) with the major in vivo phosphorylated NS5A peptides. Site-directed
      mutagenesis of Ser(2321) suggested that phosphorylation of NS5A is dispensable
      for previously described interactions with NS4A and PKR, a cellular, antiviral
      kinase that does not appear to catalyze NS5A phosphorylation. The proline-rich
      nature of the amino acid sequence flanking Ser(2321) (PLPPPRS(2321) PPVPPPR)
      suggests that a proline-directed kinase is responsible for the majority of HCV
      NS5A phosphorylation, consistent with previous kinase inhibitor studies.
FAU - Reed, K E
AU  - Reed KE
AD  - Department of Molecular Microbiology, Washington University School of Medicine,
      St. Louis, Missouri 63110-1093, USA.
FAU - Rice, C M
AU  - Rice CM
LA  - eng
GR  - CA57973/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Peptide Fragments)
RN  - 0 (Peptides)
RN  - 0 (Phosphopeptides)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Viral Nonstructural Proteins)
RN  - 452VLY9402 (Serine)
RN  - EC 2.7.11.1 (Protein Serine-Threonine Kinases)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
RN  - EC 2.7.7.48 (NS-5 protein, hepatitis C virus)
RN  - EC 2.7.7.48 (RNA-Dependent RNA Polymerase)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Animals
MH  - Cell Line
MH  - Consensus Sequence
MH  - Cricetinae
MH  - Hepacivirus/*metabolism
MH  - Mitogen-Activated Protein Kinases/metabolism
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Peptide Fragments/chemistry/isolation & purification
MH  - Peptides/chemical synthesis/chemistry/metabolism
MH  - Phosphopeptides/chemistry/isolation & purification
MH  - Phosphorylation
MH  - Protein Serine-Threonine Kinases/metabolism
MH  - RNA-Dependent RNA Polymerase/metabolism
MH  - Recombinant Fusion Proteins/chemistry/metabolism
MH  - Sequence Alignment
MH  - Sequence Homology, Amino Acid
MH  - *Serine
MH  - Transfection
MH  - Viral Nonstructural Proteins/*chemistry/*metabolism
EDAT- 1999/09/17 00:00
MHDA- 1999/09/17 00:01
CRDT- 1999/09/17 00:00
PHST- 1999/09/17 00:00 [pubmed]
PHST- 1999/09/17 00:01 [medline]
PHST- 1999/09/17 00:00 [entrez]
AID - 10.1074/jbc.274.39.28011 [doi]
AID - S0021-9258(19)52215-3 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Sep 24;274(39):28011-8. doi: 10.1074/jbc.274.39.28011.