PMID- 10383393
OWN - NLM
STAT- MEDLINE
DCOM- 19990727
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 27
DP  - 1999 Jul 2
TI  - Regulation of Hsp27 oligomerization, chaperone function, and protective activity 
      against oxidative stress/tumor necrosis factor alpha by phosphorylation.
PG  - 18947-56
AB  - The small heat shock proteins (sHsps) from human (Hsp27) and mouse (Hsp25) form
      large oligomers which can act as molecular chaperones in vitro and protect cells 
      from heat shock and oxidative stress when overexpressed. In addition, mammalian
      sHsps are rapidly phosphorylated by MAPKAP kinase 2/3 at two or three serine
      residues in response to various extracellular stresses. Here we analyze the
      effect of sHsp phosphorylation on its quaternary structure, chaperone function,
      and protection against oxidative stress. We show that in vitro phosphorylation of
      recombinant sHsp as well as molecular mimicry of Hsp27 phosphorylation lead to a 
      significant decrease of the oligomeric size. We demonstrate that both
      phosphorylated sHsps and the triple mutant Hsp27-S15D,S78D,S82D show
      significantly decreased abilities to act as molecular chaperones suppressing
      thermal denaturation and facilitating refolding of citrate synthase in vitro. In 
      parallel, Hsp27 and its mutants were analyzed for their ability to confer
      resistance against oxidative stress when overexpressed in L929 and 13.S.1.24
      cells. While wild type Hsp27 confers resistance, the triple mutant S15D,S78D,S82D
      cannot protect against oxidative stress effectively. These data indicate that
      large oligomers of sHsps are necessary for chaperone action and resistance
      against oxidative stress whereas phosphorylation down-regulates these activities 
      by dissociation of sHsp complexes to tetramers.
FAU - Rogalla, T
AU  - Rogalla T
AD  - Max-Delbruck-Centrum fur Molecule Medizin, Robert-Rossle-Str. 10, D-13122 Berlin,
      Germany.
FAU - Ehrnsperger, M
AU  - Ehrnsperger M
FAU - Preville, X
AU  - Preville X
FAU - Kotlyarov, A
AU  - Kotlyarov A
FAU - Lutsch, G
AU  - Lutsch G
FAU - Ducasse, C
AU  - Ducasse C
FAU - Paul, C
AU  - Paul C
FAU - Wieske, M
AU  - Wieske M
FAU - Arrigo, A P
AU  - Arrigo AP
FAU - Buchner, J
AU  - Buchner J
FAU - Gaestel, M
AU  - Gaestel M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (HSP27 Heat-Shock Proteins)
RN  - 0 (HSPB1 protein, human)
RN  - 0 (Heat-Shock Proteins)
RN  - 0 (Hsbp1 protein, mouse)
RN  - 0 (Hspb1 protein, rat)
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (Molecular Chaperones)
RN  - 0 (Neoplasm Proteins)
RN  - 0 (Polymers)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 12001-79-5 (Vitamin K)
RN  - 452VLY9402 (Serine)
RN  - EC 2.7.1.- (MAP-kinase-activated kinase 2)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
SB  - IM
MH  - Amino Acid Substitution
MH  - Animals
MH  - Cell Line
MH  - Circular Dichroism
MH  - Fibroblasts/metabolism
MH  - HSP27 Heat-Shock Proteins
MH  - Heat-Shock Proteins/*physiology
MH  - Humans
MH  - Intracellular Signaling Peptides and Proteins
MH  - Mice
MH  - Molecular Chaperones
MH  - Molecular Mimicry
MH  - Mutagenesis, Site-Directed
MH  - Neoplasm Proteins/genetics/*physiology
MH  - *Oxidative Stress
MH  - Phosphorylation
MH  - Polymers/metabolism
MH  - Protein Conformation
MH  - Protein-Serine-Threonine Kinases/metabolism
MH  - Rats
MH  - Serine/metabolism
MH  - Tumor Necrosis Factor-alpha/*metabolism
MH  - Vitamin K/pharmacology
EDAT- 1999/06/26 00:00
MHDA- 1999/06/26 00:01
CRDT- 1999/06/26 00:00
PHST- 1999/06/26 00:00 [pubmed]
PHST- 1999/06/26 00:01 [medline]
PHST- 1999/06/26 00:00 [entrez]
AID - 10.1074/jbc.274.27.18947 [doi]
AID - S0021-9258(19)74105-2 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Jul 2;274(27):18947-56. doi: 10.1074/jbc.274.27.18947.