Differential expression of heat shock protein (HSP70) mRNAs in rat cells.
Heat shock treatment in PC12 cells induces the synthesis of two HSP70 transcripts of 2.55 and 3.05 kb in size, while only a 70-kDa protein is shown by SDS-PAGE. Using reverse transcription-polymerase chain reaction strategy, two mRNA species were amplified from heat-shocked PC12 cells and sequenced. Both cDNAs revealed the same open reading frame encoding a single predicted 641-amino-acid polypeptide of about 70 kDa. The two 3' untranslated regions of the two mRNAs were completely different in both length and composition. Expression of the mRNA of the two corresponding genes and that of another rat HSP70 family member was investigated in PC12, Rat1 cells, and lung fibroblasts. Northern blot analysis revealed that the 2.55- and the 3.05-kb related gene transcripts are differentially expressed in the rat cell lines tested, while the third member of the subfamily is not induced. The single-copy nature of the three genes is also confirmed by Southern blot analysis.
Angeletti
B
B
Dipartimento di Medicina Sperimentale, Università de L'Aquila, Italy.
Pascale
E
E
Verna
R
R
Passarelli
F
F
Butler
R H
RH
D'Ambrosio
E
E
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis; D018076:DNA, Complementary / Q000235:genetics; D005347:Fibroblasts; D018628:Gene Dosage; D005786:Gene Expression Regulation / Q000502:physiology*; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis / Q000235:genetics*; D018869:Heat-Shock Response / Q000235:genetics; D008168:Lung / Q000166:cytology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016716:PC12 Cells; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D051381:Rats; D011916:Rats, Inbred F344; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D018840:HSP70 Heat-Shock Proteins; D012333:RNA, Messenger; D004247:DNA