The cDNA-derived amino acid sequence of chick heat shock protein Mr 90,000 (HSP 90) reveals A “DNA like” structure: Potential site of interaction with steroid receptors

We report the isolation and characterization of a cDNA clone encoding HSP47, a transformation-sensitive heat shock protein that binds to collagen. A cDNA library was prepared from total RNA isolated from heat-shocked chicken embryo fibroblasts and screened by using oligonucleotide mixtures prepared on the basis of the N-terminal amino acid sequence of biochemically purified HSP47. The cDNA insert contained 3,278 bp, which encoded a 15-amino-acid signal peptide and a mature protein coding region consisting of 390 amino acid residues; it also included part of the 5' noncoding region and a long 3' noncoding region. The deduced amino acid sequence revealed an RDEL sequence at the C terminus, which is a variant of the KDEL retention signal for retention of proteins in the endoplasmic reticulum. Northern (RNA) blot analyses and nuclear run-on assays established that the induction of HSP47 by heat shock and its suppression after transformation of chicken embryo fibroblasts by Rous sarcoma virus are regulated at the transcriptional level. A homology search revealed that this protein belongs to the serpin family, the superfamily of plasma serine protease inhibitors. Although structurally homologous to the serpins, HSP47 lacks the active site thought to be essential for the inhibition of proteases and does not appear to bind to intracellular proteases. HSP47 is the first heat shock protein found to be a member of the serpin superfamily. Conversely, it is the first serpin family member that is not secreted from cells, which could be explained by acquisition of the RDEL retention signal during evolution.

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Two-step purification and N-terminal amino acid sequence analysis of the rat Mr 90,000 heat shock protein

Analytical Biochemistry ◽

10.1016/0003-2697(88)90207-2 ◽

1988 ◽

Vol 173 (2) ◽

pp. 405-411 ◽

Cited By ~ 26

Author(s):

Marc Denis

Keyword(s):

Amino Acid ◽

Sequence Analysis ◽

Heat Shock ◽

Amino Acid Sequence ◽

Heat Shock Protein ◽

Amino Acid Sequence Analysis ◽

Terminal Amino Acid ◽

Terminal Amino ◽

Terminal Amino Acid Sequence

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HSP47: a tissue-specific, transformation-sensitive, collagen-binding heat shock protein of chicken embryo fibroblasts

Molecular and Cellular Biology ◽

10.1128/mcb.11.8.4036-4044.1991 ◽

1991 ◽

Vol 11 (8) ◽

pp. 4036-4044

Author(s):

K Hirayoshi ◽

H Kudo ◽

H Takechi ◽

A Nakai ◽

A Iwamatsu ◽

...

Keyword(s):

Amino Acid ◽

Heat Shock ◽

Amino Acid Sequence ◽

Heat Shock Protein ◽

Chicken Embryo ◽

Noncoding Region ◽

Isolation And Characterization ◽

Embryo Fibroblasts ◽

Retention Signal ◽

Chicken Embryo Fibroblasts

We report the isolation and characterization of a cDNA clone encoding HSP47, a transformation-sensitive heat shock protein that binds to collagen. A cDNA library was prepared from total RNA isolated from heat-shocked chicken embryo fibroblasts and screened by using oligonucleotide mixtures prepared on the basis of the N-terminal amino acid sequence of biochemically purified HSP47. The cDNA insert contained 3,278 bp, which encoded a 15-amino-acid signal peptide and a mature protein coding region consisting of 390 amino acid residues; it also included part of the 5' noncoding region and a long 3' noncoding region. The deduced amino acid sequence revealed an RDEL sequence at the C terminus, which is a variant of the KDEL retention signal for retention of proteins in the endoplasmic reticulum. Northern (RNA) blot analyses and nuclear run-on assays established that the induction of HSP47 by heat shock and its suppression after transformation of chicken embryo fibroblasts by Rous sarcoma virus are regulated at the transcriptional level. A homology search revealed that this protein belongs to the serpin family, the superfamily of plasma serine protease inhibitors. Although structurally homologous to the serpins, HSP47 lacks the active site thought to be essential for the inhibition of proteases and does not appear to bind to intracellular proteases. HSP47 is the first heat shock protein found to be a member of the serpin superfamily. Conversely, it is the first serpin family member that is not secreted from cells, which could be explained by acquisition of the RDEL retention signal during evolution.

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Characterization of Gmhsp26-A, a stress gene encoding a divergent heat shock protein of soybean: heavy-metal-induced inhibition of intron processing.

Molecular and Cellular Biology ◽

10.1128/mcb.8.3.1113 ◽

1988 ◽

Vol 8 (3) ◽

pp. 1113-1122 ◽

Cited By ~ 91

Author(s):

E Czarnecka ◽

R T Nagao ◽

J L Key ◽

W B Gurley

Keyword(s):

Amino Acid ◽

Heat Shock ◽

Amino Acid Sequence ◽

Genomic Sequence ◽

Stress Protein ◽

Initiation Site ◽

Gene Encoding ◽

S1 Nuclease ◽

Soybean Seedlings ◽

Nuclease Mapping

We determined the DNA sequence and mapped the corresponding transcripts of a genomic clone containing the Gmhsp26-A gene of soybean. This gene is homologous to the previously characterized cDNA clone pCE54 (E. Czarnecka, L. Edelman, F. Schöffl, and J. L. Key, Plant Mol. Biol. 3:45-58, 1984) and is expressed in response to a wide variety of physiological stresses including heat shock (HS). S1 nuclease mapping of transcripts and a comparison of the cDNA sequence with the genomic sequence indicated the presence of a soybean seedlings with either CdCl2 or CuSO4. Analysis of the 5' termini of transcripts indicated the presence of one major and at least two minor start sites. In each case, initiation occurred 27 to 30 base pairs downstream from a TATA-like motif, and thus each initiation site appears to be promoted by the activity of a separate subpromoter. The three subpromoters are all associated with sequences showing low homology to the HS consensus element of Drosophila melanogaster HS genes and are differentially induced in response to various stresses. Within the carboxyl-terminal half of the protein, hydropathy analysis of the deduced amino acid sequence indicated a high degree of relatedness to the small HS proteins. A comparison of the primary amino acid sequence of hsp26-A with sequences of the small HS proteins suggested that this stress protein is highly diverged and may therefore be specialized for stress adaptation in soybean.

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HEAT SHOCK PROTEIN 90 (90) IN AGE-DEPENDENT CHANGES IN THE NUMBER OF FIBROBLASTS IN HUMAN SKIN

Успехи геронтологии ◽

10.34922/ae.2020.33.1.004 ◽

2020 ◽

pp. 40-45

Author(s):

А. Г. Гунин ◽

Н. Н. Голубцова ◽

Н. К. Корнилова

Keyword(s):

Heat Shock ◽

Heat Shock Protein ◽

Heat Shock Protein 90 ◽

Nuclear Antigen ◽

Positive Staining ◽

Proliferating Cells ◽

Age Related ◽

Age Dependent ◽

Human Dermis ◽

Hsp 90

Целью работы стало исследование содержания белка теплового шока 90 ( HSP 90) в фибробластах дермы человека от эмбрионального развития и до глубокой старости (от 20 нед беременности до 85 лет), а также определение значения HSP 90 для возрастных изменений численности фибробластов в дерме человека. HSP 90, ядерный антиген пролиферирующих клеток ( PCNA ) выявляли в срезах кожи непрямым иммуногистохимическим методом. Результаты показали, что в коже человека от 20 нед беременности до 20 лет доля фибробластов дермы с положительной окраской на HSP 90 остается постоянной. С 21 года до 60 лет наблюдают планомерное уменьшение доли фибробластов дермы, имеющих положительную окраску на HSP 90. У людей 61-85 лет происходит резкое увеличение доли фибробластов дермы с положительной окраской на HSP 90. Возрастные изменения содержания HSP 90 положительных фибробластов в дерме статистически не связаны с возрастным уменьшением общего количества и доли PCNA -положительных фибробластов в дерме. The aim of this work was to examine the content of heat shock protein 90 ( HSP 90) in ﬁbroblasts of human dermis from the development until deep aging (from 20 weeks of pregnancy until 85 years old), and deﬁning of a role of HSP 90 in age-dependent changes in the number of ﬁbroblasts in the dermis. HSP 90, proliferating cells nuclear antigen ( PCNA ) were detected with indirect immunohistochemical technique. Results showed that a portion of ﬁbroblasts with positive staining for HSP 90 in the dermis is not changed from 20 weeks of development to 20 years old. Percent of HSP 90 positive ﬁbroblasts in dermis is decreased from 21 to 60 years old. From 61 year, the number of HSP 90 positive ﬁbroblasts in dermis is increased. Age-related changes in the number of HSP 90 positive ﬁbroblasts is not statistically associated with an age-related decrease in a total number and percent of PCNA positive ﬁbroblasts the dermis.

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Characterization of Gmhsp26-A, a stress gene encoding a divergent heat shock protein of soybean: heavy-metal-induced inhibition of intron processing

Molecular and Cellular Biology ◽

10.1128/mcb.8.3.1113-1122.1988 ◽

1988 ◽

Vol 8 (3) ◽

pp. 1113-1122

Author(s):

E Czarnecka ◽

R T Nagao ◽

J L Key ◽

W B Gurley

Keyword(s):

Amino Acid ◽

Heat Shock ◽

Amino Acid Sequence ◽

Genomic Sequence ◽

Stress Protein ◽

Initiation Site ◽

Gene Encoding ◽

S1 Nuclease ◽

Soybean Seedlings ◽

Nuclease Mapping

We determined the DNA sequence and mapped the corresponding transcripts of a genomic clone containing the Gmhsp26-A gene of soybean. This gene is homologous to the previously characterized cDNA clone pCE54 (E. Czarnecka, L. Edelman, F. Schöffl, and J. L. Key, Plant Mol. Biol. 3:45-58, 1984) and is expressed in response to a wide variety of physiological stresses including heat shock (HS). S1 nuclease mapping of transcripts and a comparison of the cDNA sequence with the genomic sequence indicated the presence of a soybean seedlings with either CdCl2 or CuSO4. Analysis of the 5' termini of transcripts indicated the presence of one major and at least two minor start sites. In each case, initiation occurred 27 to 30 base pairs downstream from a TATA-like motif, and thus each initiation site appears to be promoted by the activity of a separate subpromoter. The three subpromoters are all associated with sequences showing low homology to the HS consensus element of Drosophila melanogaster HS genes and are differentially induced in response to various stresses. Within the carboxyl-terminal half of the protein, hydropathy analysis of the deduced amino acid sequence indicated a high degree of relatedness to the small HS proteins. A comparison of the primary amino acid sequence of hsp26-A with sequences of the small HS proteins suggested that this stress protein is highly diverged and may therefore be specialized for stress adaptation in soybean.

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