A nuclear protein associated with lethal heat shock of HL-60 cells.
The responses to stress in living cells are well known. Thermal stress causes decreased protein synthesis as well as rapid induction of heat shock proteins (hsps), or alternately termed stress proteins (sps). The exposure of cultured promyelocytic leukemia cells (HL-60) to a 45 degrees C lethal heat shock for 1 h elicited synthesis and phosphorylation of a polypeptide M(r) 48,000 and pI 7.5 (p 48) as visualized by two-dimensional polyacrylamide gel ultra-microelectrophoresis. p 48, which was not observed at sublethal temperatures (39 and 41 degrees C), was synthesized during all phases of the cell cycle but was phosphorylated only in G0 + G1 and S-phases. The appearance of p 48 was marked by a concomitant and reciprocal reduction in hsps or sps 70 and 90. Distinct protease V8 fragment maps of p 48, hsps 70 and 90 in conjunction with immunochemical determination indicated vast differences in their primary structures. These facts suggest that p 48 was not formed from coalesced breakdown products of hsps 70 or 90. Western blotting showed that p 48 possessed the same immunochemical determinants as two other proteins with the same molecular mass but different isoelectric points. In an association assay, p 48 was shown to bind with actins and hsp 90 from HL-60 nuclei.
Pipkin
J L
JL
Division of Genetic Toxicology, Food and Drug Administration, National Center for Toxicological Research, Jefferson, Arkansas 72079.
Hinson
W G
WG
Lyn-Cook
L E
LE
Burns
E R
ER
Sheehan
D
D
Casciano
D A
DA
D002453:Cell Cycle; D016923:Cell Death; D002455:Cell Division; D002460:Cell Line; D002467:Cell Nucleus / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D006358:Hot Temperature; D006801:Humans; D007700:Kinetics; D015473:Leukemia, Promyelocytic, Acute; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D013462:Sulfur Radioisotopes; D013696:Temperature
D006360:Heat-Shock Proteins; D009687:Nuclear Proteins; D013462:Sulfur Radioisotopes; D008715:Methionine