Methylations of 70,000-Da heat shock proteins in 3T3 cells: alterations by arsenite treatment, by different stages of growth and by virus transformation.
We have characterized the basic amino acid methylation of three members of the 70,000-Da heat shock protein superfamily, hsp68, hsc70, and BiP, in Balb/c 3T3 cells. It appears that a lysyl residue is the only methylation site in BiP and that both lysyl and arginyl residues are methylated in hsp68 and hsc70. In all cases, epsilon-N-trimethyllysine is the predominant methyllysine species. Both NG-monomethylarginine and NG,NG-dimethylarginine are identified as the methylarginine species. The stoichiometry of the methylation is indirectly determined by using the amount of actin methylation as a reference. Three, four, and four methyl groups are incorporated into lysyl residues of hsp68, hsc70, and BiP, respectively. The level of lysyl methylation in hsc70 remains unchanged under different growth conditions. On the other hand, the arginyl methylation in hsc70 varies considerably. In confluent Balb/c 3T3 cells, there are 1.8 and 1.3 methyl groups in dimethylarginine and monomethyl-arginine, respectively. In nonconfluent cells, the amount of monomethylarginine is similar to that in confluent cells, but dimethylarginine is not detectable. Furthermore, in both confluent and nonconfluent cells, the level of monomethylarginine is reduced 5- to 10-fold after arsenite treatment. However, in 3T3 cells transformed by Rous sarcoma virus (SR-RSV 3T3 cells), the level of arginine methylation is constitutively lower and cannot be reduced further by arsenite.
Wang
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Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.
Lin
J M
JM
Lazarides
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D016475:3T3 Cells; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000378:metabolism; D001151:Arsenic / Q000494:pharmacology*; D018053:Arsenites; D001358:Avian Sarcoma Viruses / Q000235:genetics*; D002455:Cell Division; D002472:Cell Transformation, Viral; D002852:Chromatography, Ion Exchange; D015180:Electrophoresis, Gel, Two-Dimensional; D006360:Heat-Shock Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008239:Lysine / Q000031:analogs & derivatives / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D008745:Methylation; D051379:Mice; D008970:Molecular Weight; D019323:omega-N-Methylarginine
D018053:Arsenites; D006360:Heat-Shock Proteins; D019323:omega-N-Methylarginine; C003712:trimethyllysine; C018524:N,N-dimethylarginine; D001120:Arginine; D008715:Methionine; D008239:Lysine; C015001:arsenite; D001151:Arsenic