PMID- 10362514
OWN - NLM
STAT- MEDLINE
DCOM- 19990628
LR  - 20091119
IS  - 0006-291X (Print)
IS  - 0006-291X (Linking)
VI  - 259
IP  - 2
DP  - 1999 Jun 7
TI  - Identification of phosphorylation sites in the PKD1-encoded protein C-terminal
      domain.
PG  - 356-63
AB  - The PKD1-encoded protein, "polycystin-1", has a large N-terminal extracellular
      portion, multiple transmembrane domains, and a short intracellular C-terminal
      tail with four tyrosine residues and two putative sites for serine
      phosphorylation. Its function in kidney development and autosomal dominant
      polycystic kidney disease (ADPKD) is still unknown. We have subcloned the cDNA
      encoding the polycystin-1 C-terminal domain (PKD1-CTD) into a prokaryotic
      expression vector, and site-directed mutagenesis was performed to target the four
      tyrosine residues and four serine residues in two putative phosphorylation sites.
      In vitro phosphorylation assays were conducted on both wild type and mutant
      PKD1-CTD fusion proteins. It was found that the wild type PKD1-CTD and all mutant
      fusion proteins, except S4251G/S4252G, could be phosphorylated by lysates from
      cultured normal human renal collecting tubule (NHCT) cells, as well as by
      commercially purified cAMP-dependent protein kinase (PKA). The phosphorylation of
      the PKD1-CTD fusion protein by NHCT lysates was greatly enhanced by cAMP and its 
      analog 8-Br-cAMP, and inhibited by the specific PKA inhibitors PKI(6-22) and
      H-89. Activators and inhibitors of protein kinase C (PKC) had no effects on the
      phosphorylation of the PKD1-CTD fusion protein. Using commercially purified
      pp60(c-src) (c-src) it was also shown that the PKD1-CTD fusion protein could be
      phosphorylated by c-src in vitro, and that this phosphorylation could be
      abolished by a mutation Y4237F. By comparing the amino acid sequence at 4249-4253
      (RRSSR) with the consensus sequence for PKA phosphorylation (RRXSX), we suggest
      that the serine residue at 4252 is the target of phosphorylation by a
      cAMP-dependent protein kinase in NHCT cell lysates. In addition, we suggest that 
      Y4237 might be phosphorylated by c-src in living cells.
CI  - Copyright 1999 Academic Press.
FAU - Li, H P
AU  - Li HP
AD  - Department of Medicine, Mount Sinai School of Medicine, New York, New York 10029,
      USA. Hsi-Ping_Li@mtplink.mssm.edu
FAU - Geng, L
AU  - Geng L
FAU - Burrow, C R
AU  - Burrow CR
FAU - Wilson, P D
AU  - Wilson PD
LA  - eng
GR  - R01 DK448833/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochem Biophys Res Commun
JT  - Biochemical and biophysical research communications
JID - 0372516
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Proteins)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (TRPP Cation Channels)
RN  - 0 (polycystic kidney disease 1 protein)
RN  - EC 2.7.10.2 (Proto-Oncogene Proteins pp60(c-src))
RN  - EC 2.7.11.11 (Cyclic AMP-Dependent Protein Kinases)
SB  - IM
MH  - Cells, Cultured
MH  - Cyclic AMP-Dependent Protein Kinases/metabolism
MH  - Enzyme Activation/drug effects
MH  - Enzyme Inhibitors/pharmacology
MH  - Humans
MH  - Kidney Tubules/metabolism
MH  - Phosphorylation
MH  - Polycystic Kidney, Autosomal Dominant/genetics
MH  - Proteins/*chemistry
MH  - Proto-Oncogene Proteins pp60(c-src)/metabolism
MH  - Recombinant Fusion Proteins/chemistry
MH  - TRPP Cation Channels
EDAT- 1999/06/11 00:00
MHDA- 1999/06/11 00:01
CRDT- 1999/06/11 00:00
PHST- 1999/06/11 00:00 [pubmed]
PHST- 1999/06/11 00:01 [medline]
PHST- 1999/06/11 00:00 [entrez]
AID - S0006-291X(99)90780-5 [pii]
AID - 10.1006/bbrc.1999.0780 [doi]
PST - ppublish
SO  - Biochem Biophys Res Commun. 1999 Jun 7;259(2):356-63. doi:
      10.1006/bbrc.1999.0780.