Structure and Expression of the Human Gene Encoding Major Heat Shock Protein HSP70

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 1985 Feb 1

PMID 2858050

Citations 161

Authors

B Wu

C Hunt

R Morimoto

Affiliations

Soon will be listed here.

Abstract

We have cloned a human gene encoding the 70,000-dalton heat shock protein (HSP70) from a human genomic library, using the Drosophila HSP70 gene as a heterologous hybridization probe. The human recombinant clone hybridized to a 2.6-kilobase polyadenylated mRNA from HeLa cells exposed to 43 degrees C for 2 h. The 2.6-kilobase mRNA was shown to direct the translation in vitro of a 70,000-dalton protein similar in electrophoretic mobility to the HSP70 synthesized in vivo. From the analysis of S1 nuclease-resistant mRNA-DNA hybrids, the HSP70 gene appears to be transcribed as an uninterrupted mRNA of 2.3 kilobases. We show that the cloned HSP70 gene contains the sequences necessary for heat shock-induced expression by two criteria. First, hamster cells transfected with a subclone containing the HSP70 gene and flanking sequences synthesized a HSP70-like protein upon heat shock. Second, human cells transfected with a chimeric gene containing the 5' flanking sequences of the HSP70 gene and the coding sequences of the bacterial chloramphenicol acetyltransferase gene transcribed the chimeric gene upon heat shock. We show that the HSP70 mRNA transcribed in an adenovirus 5 transformed human cell line (293 cells) is identical to the HSP70 mRNA induced by heat shock.

Citing Articles

Heat application in live cell imaging.

Sistemich L, Ebbinghaus S FEBS Open Bio. 2024; 14(12):1940-1954.

PMID: 39489617 PMC: 11609584. DOI: 10.1002/2211-5463.13912.

The dance of proteostasis and metabolism: Unveiling the caloristatic controlling switch.

Schroeder H, de Lemos Muller C, Heck T, Krause M, Homem de Bittencourt Jr P Cell Stress Chaperones. 2024; 29(1):175-200.

PMID: 38331164 PMC: 10939077. DOI: 10.1016/j.cstres.2024.02.002.

Role of Heat Shock Factors in Stress-Induced Transcription: An Update.

Bunch H, Calderwood S Methods Mol Biol. 2023; 2693:25-38.

PMID: 37540424 DOI: 10.1007/978-1-0716-3342-7_3.

Developing a Temperature-Inducible Transcriptional Rheostat in Neurospora crassa.

Tabilo-Agurto C, Del Rio-Pinilla V, Eltit-Villarroel V, Goity A, Munoz-Guzman F, Larrondo L mBio. 2023; 14(1):e0329122.

PMID: 36744948 PMC: 9973361. DOI: 10.1128/mbio.03291-22.

Single-Cell Sequencing to Identify Six Heat Shock Protein (HSP) Genes-Mediated Progression Subtypes of Clear Cell Renal Cell Carcinoma.

Li Q, Lu M, Zhang Z, Zhang R Int J Gen Med. 2021; 14:3761-3773.

PMID: 34326662 PMC: 8315815. DOI: 10.2147/IJGM.S318271.

References

Maxam A, Gilbert W . Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980; 65(1):499-560. DOI: 10.1016/s0076-6879(80)65059-9. View

Shen C, Maniatis T . The organization of repetitive sequences in a cluster of rabbit beta-like globin genes. Cell. 1980; 19(2):379-91. DOI: 10.1016/0092-8674(80)90512-7. View

Fritsch E, Lawn R, Maniatis T . Molecular cloning and characterization of the human beta-like globin gene cluster. Cell. 1980; 19(4):959-72. DOI: 10.1016/0092-8674(80)90087-2. View

Cleveland D, Lopata M, MacDonald R, Cowan N, Rutter W, Kirschner M . Number and evolutionary conservation of alpha- and beta-tubulin and cytoplasmic beta- and gamma-actin genes using specific cloned cDNA probes. Cell. 1980; 20(1):95-105. DOI: 10.1016/0092-8674(80)90238-x. View

Lamb R, Lai C . Sequence of interrupted and uninterrupted mRNAs and cloned DNA coding for the two overlapping nonstructural proteins of influenza virus. Cell. 1980; 21(2):475-85. DOI: 10.1016/0092-8674(80)90484-5. View

Drouin J . Cloning of human mitochondrial DNA in Escherichia coli. J Mol Biol. 1980; 140(1):15-34. DOI: 10.1016/0022-2836(80)90354-x. View

Nagata S, Mantei N, Weissmann C . The structure of one of the eight or more distinct chromosomal genes for human interferon-alpha. Nature. 1980; 287(5781):401-8. DOI: 10.1038/287401a0. View

Bell G, Pictet R, Rutter W . Analysis of the regions flanking the human insulin gene and sequence of an Alu family member. Nucleic Acids Res. 1980; 8(18):4091-109. PMC: 324221. DOI: 10.1093/nar/8.18.4091. View

Thomas P . Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980; 77(9):5201-5. PMC: 350025. DOI: 10.1073/pnas.77.9.5201. View

10.

Kioussis D, Eiferman F, van de Rijn P, Gorin M, Ingram R, Tilghman S . The evolution of alpha-fetoprotein and albumin. II. The structures of the alpha-fetoprotein and albumin genes in the mouse. J Biol Chem. 1981; 256(4):1960-7. View

11.

Ryffel G, Muellener D, Wyler T, Wahli W, Weber R . Transcription of single-copy vitellogenin gene of Xenopus involves expression of middle repetitive DNA. Nature. 1981; 291(5814):429-31. DOI: 10.1038/291429a0. View

12.

Wang C, Gomer R, Lazarides E . Heat shock proteins are methylated in avian and mammalian cells. Proc Natl Acad Sci U S A. 1981; 78(6):3531-5. PMC: 319603. DOI: 10.1073/pnas.78.6.3531. View

13.

Slater A, Cato A, Sillar G, Kioussis J, BURDON R . The pattern of protein synthesis induced by heat shock of HeLa cells. Eur J Biochem. 1981; 117(2):341-6. DOI: 10.1111/j.1432-1033.1981.tb06343.x. View

14.

Hentschel C, Birnstiel M . The organization and expression of histone gene families. Cell. 1981; 25(2):301-13. DOI: 10.1016/0092-8674(81)90048-9. View

15.

Lis J, Ish-Horowicz D, Pinchin S . Genomic organization and transcription of the alpha beta heat shock DNA in Drosophila melanogaster. Nucleic Acids Res. 1981; 9(20):5297-310. PMC: 327521. DOI: 10.1093/nar/9.20.5297. View

16.

Deininger P, Jolly D, Rubin C, Friedmann T, Schmid C . Base sequence studies of 300 nucleotide renatured repeated human DNA clones. J Mol Biol. 1981; 151(1):17-33. DOI: 10.1016/0022-2836(81)90219-9. View

17.

Jagadeeswaran P, Forget B, Weissman S . Short interspersed repetitive DNA elements in eucaryotes: transposable DNA elements generated by reverse transcription of RNA pol III transcripts?. Cell. 1981; 26(2 Pt 2):141-2. DOI: 10.1016/0092-8674(81)90296-8. View

18.

Singer M . SINEs and LINEs: highly repeated short and long interspersed sequences in mammalian genomes. Cell. 1982; 28(3):433-4. DOI: 10.1016/0092-8674(82)90194-5. View

19.

Ingolia T, Craig E . Drosophila gene related to the major heat shock-induced gene is transcribed at normal temperatures and not induced by heat shock. Proc Natl Acad Sci U S A. 1982; 79(2):525-9. PMC: 345776. DOI: 10.1073/pnas.79.2.525. View

20.

Schmid C, Jelinek W . The Alu family of dispersed repetitive sequences. Science. 1982; 216(4550):1065-70. DOI: 10.1126/science.6281889. View