Members Of HSP70 Gene Family Research Articles

Identification of HSP70 gene family members in Fritillaria cirrhosa and expression analysis in different tissues under high temperature stress

The heat shock protein 70 family contains the stress proteins ubiquitous in plants. These proteins are involved in the responses to different abiotic stress conditions and have highly conserved gene sequences. However, little is known about the molecular mechanisms of Fritillaria cirrhosa in response to high-temperature stress. Here, 26 HSP70s, FcHSP70-1 to FcHSP70-26, were identified from the transcriptome data of root, bulb, stem, leaf, and fruit samples of F. cirrhosa. The proteins encoded by FcHSP70s had the lengths ranging from 560 aa to 944 aa, with the molecular weight of 61.64-100.01 kDa and the theoretical isoelectric point between 5.00 and 6.59. The secondary structural elements of HSP70s were mainly random coils and α-helixes. Subcellular localization prediction revealed that FcHSP70s were distributed in mitochondria, chloroplasts, nuclei, endoplasmic reticulum, and cytoplasm. The phylogenetic tree showed that 7 members of the HSP70 family belonged to the Dnak subfamily and 19 members belonged to the HSP110/SSE subfamily. In addition, the qRT-PCR results showed that the expression of FcHSP70-5, FcHSP70-8, FcHSP70-17, FcHSP70-18, and FcHSP70-23 in F. cirrhosa was significantly up-regulated at 35 ℃, which indicated that these genes might play a role in the response to high temperature stress. In addition, compared with other tissues, stems and leaves were sensitive to high temperature stress, with the expression of 18 genes up-regulated by 18.18 and 8.03 folds on average, respectively. These findings provide valuable information about the molecular mechanism of HSP70s of F. cirrhosa in response to high temperature stress.

Identification and expression analysis of the HSP70 gene family under abiotic stresses in Litchi chinensis

HSP70 protein, as an important member of the heat shock protein (HSP) family, plays an important role in plant growth, development, and response to biotic and abiotic stresses. In order to explore the role of HSP70 gene family members in Litchi chinensis under low temperature, high temperature, drought, and salt stress, bioinformatics methods were used to identify the HSP70 gene family members within the entire L. chinensis genome. The expression of these genes under various abiotic stresses was then detected using quantitative real-time PCR (qRT-PCR). The results showed that the LcHSP70 gene family consisted of 18 members, which were unevenly distributed across ten L. chinensis chromosomes. The LcHSP70 protein contained 479-851 amino acids, with isoelectric points ranging from 5.07 to 6.95, and molecular weights from 52.44 kDa to 94.07 kDa. The predicted subcellular localization showed that LcHSP70 protein was present in the nucleus, cytoplasm, endoplasmic reticulum, mitochondria, and chloroplast. Phylogenetic analysis divided the LcHSP70 proteins into five subgroups, namely Ⅰ, Ⅱ, Ⅲ, Ⅳ, and Ⅵ. The promoter regions of the LcHSP70 genes contained various cis-acting elements related to plant growth, development, hormone response, and stress response. Moreover, the expression of LcHSP70 genes displayed distint tissue-specific expression level, categorized into universal expression and specific expression. From the selected 6 LcHSP70 genes (i.e., LcHSP70-1, LcHSP70-5, LcHSP70-10, LcHSP70-14, LcHSP70-16, and LcHSP70-18), their relative expression levels were assessed under different abiotic stresses using qRT-PCR. The results indicated that the gene family members exhibited diverse responses to low temperature, high temperature, drought, and salt stress, with significant variations in their expression levels across different time periods. These results provide a foundation for further exploration of the function of the LcHSP70 gene family.

HSP70 Gene Family in Brassica rapa: Genome-Wide Identification, Characterization, and Expression Patterns in Response to Heat and Cold Stress.

Heat shock proteins protect plants from abiotic stress, such as salt, drought, heat, and cold stress. HSP70 is one of the major members of the heat shock protein family. To explore the mechanism of HSP70 in Brassica rapa, we identified 28 putative HSP70 gene family members using state-of-the-art bioinformatics-based tools and methods. Based on chromosomal mapping, HSP70 genes were the most differentially distributed on chromosome A03 and the least distributed on chromosome A05. Ka/Ks analysis revealed that B. rapa evolution was subjected to intense purifying selection of the HSP70 gene family. RNA-sequencing data and expression profiling showed that heat and cold stress induced HSP70 genes. The qRT-PCR results verified that the HSP70 genes in Chinese cabbage (Brassica rapa ssp. pekinensis) are stress-inducible under both cold and heat stress. The upregulated expression pattern of these genes indicated the potential of HSP70 to mitigate environmental stress. These findings further explain the molecular mechanism underlying the responses of HSP70 to heat and cold stress.

Delineation of temperature-humidity index (THI) as indicator of heat stress in riverine buffaloes (Bubalus bubalis) of a sub-tropical Indian region.

The erstwhile developed temperature-humidity index (THI) has been popularly used to indicate heat stress in dairy cattle and often in buffaloes. However, scientific literature suggests differences in thermotolerance and physiological responses to heat stress between cattle and buffalo. Therefore, THI range used to indicate degree of heat stress (mild, moderate, and severe) in cattle should be recalibrated for indicating heat stress in buffaloes. The present study was carried out to delineate THI range to indicate onset and severity of heat stress in buffaloes based on physiological, biochemical, and expression profiling of heat shock response (HSR) genes in animals at different THI. The result indicated early onset of heat stress in buffaloes as compared to cattle. Physiological and biochemical parameters indicated onset of mild signs of heat stress in buffaloes at THI 68-69. Significant deviation in these parameters was again observed at THI range 73-76. At THI 77-80, the physiological and biochemical responses of animals were further intensified indicating extreme alteration in homeostasis. The in vivo expression profiling of HSR genes indicated that members of Hsp70 gene family are expressed in a temporal pattern over different THIs, whereas expressions of Hsf genes were evident during intense heat stress. Overall, the study established that amplitude of heat shock response and THI range for indicating severity of thermal stress for buffaloes are not in unison to cattle. The study also suggests skin temperature of the poll region could be used as non-invasive tool for monitoring heat stress in dairy buffaloes.

In Silico Analysis of HSP70 Gene Family in Bovine Genome

Heat shock proteins (HSPs), members of molecular chaperones families fulfill essential roles under normal conditions and provide protection and adaptation during and after stress. Among different HSPs, HSP70 kDa family of proteins is most abundant and well-studied in human and mouse but has not yet been characterized in bovines. In silico analysis was performed to characterize members of HSP70 gene family in bovine genome and a total of 17 genes of bovine HSP70 gene family were identified. The members of HSP70 family were distributed over 12 chromosomes with gene size ranging from 1911 (HSPA2) to 54,017bp (HSPA4). Five genes were intronless, while rest of 12 genes were multiexonic. Phylogenetic analysis of HSP70 gene family distinguished them into eight major evolutionary groups wherein members of group 1 were most divergent and quite dissimilar than from rest of the HSP70 sequences. Domain structure of all bovine HSP70 genes was conserved and three signature patterns HSP70_1, HSP70_2, and HSP70_3 were identified. HSPA8, HSP9, and HSPA1A showed comparatively higher expression in majority of tissues. Like humans, bovine HSP70 family was characterized by remarkable evolutionary diversity. The analysis also suggested resemblance of bovine HSP70 family to that of human compared to mouse. Overall, the study indicates the presence of diversity for structure, function, localization, and expression in the bovine HSP70 family chaperons which could form the basis to understand thermotolerance/adaptive changes in the bovines.

Genome-Wide Identification of Hsp70 Genes in the Large Yellow Croaker (Larimichthys crocea) and Their Regulated Expression Under Cold and Heat Stress.

Heat shock proteins 70 (Hsp70) are required for key cellular processes and responses to environmental changes, however, there are an unknown number of hsp70 gene family members in the large yellow croaker (Larimichthys crocea). In the present study, 17 hsp70 genes were identified through the genome of the large yellow croaker. These genes are divided into seven evolutionarily distinct groups according to a phylogenetic tree. The orthologs of these hsp70 genes were found in humans and zebrafish. The expression patterns of the hsp70 gene family in the large yellow croaker under cold and heat stress were studied by examining transcriptome data. Six out of 17 genes were significantly unregulated or downregulated after cold or heat stress. There were two genes significantly upregulated and two genes downregulated in the liver after cold treatment, while after heat treatment, five genes were significantly upregulated, and no genes were significantly downregulated. Three expression patterns were detected: strictly heat-inducible hsp70, constitutively expressed and moderately heat-inducible hsp70, and constitutively expressed and less stress-dependent hsp70 genes. All the findings will contribute to a better understanding of the biological function of hsp70s in defending against thermal challenges.

HSP70 (HSPA1) polymorphisms in former workers with chronic mercury vapor exposure.

To investigate 4 loci of 3 HSP70 genes in caustic soda production plant former workers, who have been exposed to metallic mercury vapors for a long time, and including numerous cases of chronic mercury intoxication (CMI). Polymorphisms in HSP70 gene family members (HSP1A1 (+190G/C, rs1043618), HSPA1B (+1267A/G and +2074G/C, rs1061581) and HSP1AL (+2437T/C, rs2227956)) genes were studied among 120 male workers involved in caustic soda production by mercury electrolysis at 2 plants in Eastern Siberia. These subjects had been chronically exposed to metallic mercury vapors for > 5 years and divided into 3 groups based on the occurrence and time of the CMI diagnosis, or absence of this disease. The Group 1 consisted of individuals (N = 46), who had had contact with mercury but were not diagnosed with the CMI. The Group 2 included workers (N = 56), who were diagnosed with the CMI longer than 14 years ago. The Group 3 consisted of the subjects (N = 18), who had been diagnosed with the CMI 3-5 years ago. The logistic regression analysis was used for 3 genetic models with and without adjustment for age and duration of mercury vapor exposure. We found that genotypes СС-HSPA1A (+190G/C) and GG-HSPA1B (+1267A/G) had a high predictive risk of the CMI development (adjusted odds ratio (ORadj) = 5.58, p = 0.026 and ORadj = 14.7, p = 0.0015, respectively). Twelve individuals with the CMI had a specific combination of СС-HSPA1A (+190G/C) and GG-HSPA1B (+1267A/G) genotypes, which strongly associated with the diagnosis (ORadj = 12.3, p = 0.0285). Moreover, significant association with the CMI was also obtained for the haplotype G-C of 1267A/G and 190G/C polymorphisms (OR = 2.1, p = 0.018). The association of СС-HSPA1A (+190G/C) and GG-HSPA1B (+1267A/G) genotypes and their combination for the CMI individuals suggests the role for HSPA1 genes in mercury-dependent mechanisms of the CMI development and progression. Int J Occup Med Environ Health 2017;30(1):77-85.

Endothelial nitric oxide synthase induces heat shock protein HSPA6 (HSP70B′) in human arterial smooth muscle cells

Endothelial nitric oxide synthase (eNOS) is the major source of nitric oxide (NO) production in blood vessels. One of the pleitropic functions of eNOS derived NO is to inhibit vascular smooth muscle cell proliferation in the blood vessel wall, and whose dysfunction is a primary cause of atherosclerosis and restenosis. In this study there was an interest in examining the gene profile of eNOS adenoviral (Ad-eNOS) transduced human coronary artery smooth muscle cells (HCASMC) to further understand the eNOS inhibitory effect on smooth muscle cell proliferation. To this aim a whole genome wide analysis of eNOS transduced HCASMCs was performed. A total of 19 genes were up regulated, and 31 genes down regulated in Ad-eNOS transduced HCASMCs compared to cells treated with an empty adenovirus. Noticeably, a cluster of HSP70 gene family members was amongst the genes up regulated. Quantitative PCR confirmed that transcripts for HSPA1A (HSP70A), HSPA1B (HSP70B) and HSPA6 (HSP70B′) were elevated 2, 1.7 and 14-fold respectively in Ad-eNOS treated cells. The novel gene HSPA6 was further explored as a potential mediator of eNOS signaling in HCASMC. Immunoblotting showed that HSPA6 protein was induced by Ade-NOS. To functionally examine the effect of HSPA6 on SMCs, an adenovirus harboring the HSPA6 gene under the control of a constitutive promoter was generated. Transduction of HCASMCs with Ad-HSPA6 inhibited SMC proliferation at 3 and 6 days post serum growth stimulation, and paralleled the Ad-eNOS inhibition of SMC growth. The identification in this study that HSPA6 overexpression inhibits SMC proliferation coupled with the recent finding that inhibition of HSP90 has a similar effect, progresses the field of targeting HSPs for vascular repair.

Genome-wide analysis of HSP70 superfamily in Gossypium raimondii and the expression of orthologs in Gossypium hirsutum

Heat shock 70 (HSP70) proteins are highly conserved molecular chaperones widely existed in the plant kingdom which are involved in cellular protein folding process. In this study, comprehensive evolutionary analyses of the Gossypium raimondii HSP70 gene family members are conducted and 30 HSP70 genes are identified. The gene structure, chromosome distribution, gene duplication and phylogenic evolution of this family are further analyzed. The results reveal that HSP70 family genes can be clustered into several major subgroups based on their sub-cellular locations, and the gene structures are relatively conserved in each subgroup. Both tandem duplications and chromosome segmental duplications are found to contribute to the expansion of HSP70 gene family. Evolutionary analysis of HSP70s in diverse species reveals that the differentiation of HSP70 subgroups occurred before the multi-cell and single-cell differentiation, with the cytoplasmic HSP70s multiple amplified. The expression pattern of HSP70 genes under series of fiber development stages indicates that many HSP70 genes may participate in fiber development processes including fiber initiation and elongation. This study provides the complete profiles of cotton HSP70 family genes for future study on their functions related to the molecular mechanisms of fiber development.

Clonaje y caracterización molecular de dos genes homólogos hsp70 del hongo dimórficoParacoccidioides brasilensis.

Paracoccidioides brasiliensis, a dimorphic fungus, is the etiologic agent of Paracoccidioidomycosis (PCM), one of the most important systemic mycosis in Latin America. Two genes (2.2 and 1DB5) were cloned, characterized and sequenced; they showed homology with members of hsp70 gene family. By using several probe fragments derived from these genes, levels of expression for each gene were determined by Northern blot during transition to the yeast phase. The highest level of hsp70 transcript occurred between 30 min to 6 hours after temperature shift, with significant reduction after 36-48 hours. However, after 72 hours, the level of the transcription increased until yeast phase was reached. As a response to temperature increase, hsp 70 genes are expressed during the transition phase and possibly play a role in the differentiation process.

Application of modified differential display technique for cloning and sequencing of the 3' region from three putative members of wheat HSP70 gene family.

We have modified the differential display technique to isolate 3' regions from different members of the wheat HSP70 gene family. An HSP70 gene family-specific degenerate primer was used as a 5' primer in place of the arbitrary primer used in the original technique. We cloned and sequenced three cDNA fragments that were differentially expressed in heat stressed wheat seedlings. Based on the high similarity between predicted translation products of these three sequences and known members of the HSP70 family from plants, these cDNAs were identified as members of the HSP70 gene family. Two of these members appeared distinct in the 3' non-coding region with only 48% identity. Therefore differential display could successfully be used to isolate 3' regions of different members of a multigene family in a relatively short period, even if the members had highly similar protein-coding regions.

Chromosomal Localization of Five Murine HSP70 Gene Family Members: Hsp70.1, Hsp70-2, Hsp70-3, Hsc70t, and Grp78

The 70,000-D heat shock protein (HSP70) gene family includes both heat-inducible and constitutively expressed genes. We have mapped five murine HSP70 genes to specific sites on three separate chromosomes. Southern blot analysis of Chinese hamster X mouse somatic cell DNAs was used to assign the gene for the 78,000-D glucose-regulated protein (Grp78) to Chromosome (Chr) 2, the mule germ cell-specific Hsp70-2 and Hsc70t genes to Chr 12 and Chr 17, respectively, and the heat-inducible Hsp70-3 gene also to Chr 17. Southern blot analysis of DNA from the progeny of two multilocus crosses confirmed the Grp78 location on Chr 2 and suggested the order: centromere-Vim-Abl-Grp78-Hc. Similar analysis also confirmed the initial Hsp70-2 assignment to Chr 12 with the order: Hsp70-2-Aat-Igh . The Hsp70-3 and Hsc70t genes on Chr 17, along with the heat-inducible Hsp70-1 gene, were further localized by Southern blot analysis of genomic clones to the H-2 histocompatibility region with the order: Hsp70-1-Hsp70-3-Hsc70t-Bat-6 (human G7a, valyl-tRNA synthetase).

Identification and Sequence Analysis of a New Member of the Mouse HSP70 Gene Family and Characterization of Its Unique Cellular and Developmental Pattern of Expression in the Male Germ Line

A unique member of the mouse HSP70 gene family has been isolated and characterized with respect to its DNA sequence organization and expression. The gene contains extensive similarity to a heat shock-inducible HSP70 gene within the coding region but diverges in both 3' and 5' nontranslated regions. The gene does not yield transcripts in response to heat shock in mouse L cells. Rather, the gene appears to be activated uniquely in the male germ line. Analysis of RNA from different developmental stages and from enriched populations of spermatogenic cells revealed that this gene is expressed during the prophase stage of meiosis. A transcript different in size from the major heat-inducible mouse transcripts is most abundant in meiotic prophase spermatocytes and decreases in abundance in postmeiotic stages of spermatogenesis. This pattern of expression is distinct from that observed for another member of this gene family, which was previously shown to be expressed abundantly in postmeiotic germ cells. These observations suggest that specific HSP70 gene family members play distinct roles in the differentiation of the germ cell lineage in mammals.

Identification and sequence analysis of a new member of the mouse HSP70 gene family and characterization of its unique cellular and developmental pattern of expression in the male germ line.

A unique member of the mouse HSP70 gene family has been isolated and characterized with respect to its DNA sequence organization and expression. The gene contains extensive similarity to a heat shock-inducible HSP70 gene within the coding region but diverges in both 3' and 5' nontranslated regions. The gene does not yield transcripts in response to heat shock in mouse L cells. Rather, the gene appears to be activated uniquely in the male germ line. Analysis of RNA from different developmental stages and from enriched populations of spermatogenic cells revealed that this gene is expressed during the prophase stage of meiosis. A transcript different in size from the major heat-inducible mouse transcripts is most abundant in meiotic prophase spermatocytes and decreases in abundance in postmeiotic stages of spermatogenesis. This pattern of expression is distinct from that observed for another member of this gene family, which was previously shown to be expressed abundantly in postmeiotic germ cells. These observations suggest that specific HSP70 gene family members play distinct roles in the differentiation of the germ cell lineage in mammals.