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Genetic polymorphism in Hsp90AA1 gene is associated with the thermotolerance in Chinese Holstein cows.

ABSTRACT: The heat shock protein 90 (Hsp90) is a copious and ubiquitous molecular chaperone which plays an essential role in many cellular biological processes. The objective of this study was to identify single nucleotide polymorphisms (SNPs) in the Hsp90AA1 gene and to determine their association with heat stress traits in Chinese Holstein cattle breed. Direct sequencing was used to identify new SNPs. Luciferase reporter assay methods were used to assess g.-?87G?>?C and g.4172A?>?G loci in the promoter activity and 3'-UTR, respectively. Quantitative real-time PCR was utilized to quantify the gene expression profile. Five SNPs were identified in 130 multiparous lactating cows: one SNP in the promoter, three SNPs in the coding region, and one in 3'-UTR were novel and reported for the first time in this study. As a result of promoter assay using dual luciferase assay system, the genotype CC showed the highest transcription activity region (13.67?±?0.578) compared to the wild-type GG (3.24?±?0.103). On the other hand, the result revealed that one of the selected microRNAs (dme-miR-2279-5p) was found to interact with the Hsp90AA1 3'-UTR sequence and to suppress the reporter activity markedly in the presence of the allele G (2.480?±?0.136). The expression of Hsp90AA1 in cow bearing mutant allele C was higher (4.18?±?0.928) than cows bearing wild-type allele G (1.008?±?0.0.129) in stress season. In summary, there was an association between genetic variations in the Hsp90AA1 and thermoresistance. This association could be used as a marker in genetic selection for heat tolerance in Chinese Holstein cattle breeds.

SUBMITTER: Badri TM

PROVIDER: S-EPMC6045533 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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Genetic polymorphism in Hsp90AA1 gene is associated with the thermotolerance in Chinese Holstein cows.

Badri T M TM Chen K L KL Alsiddig M A MA Li Lian L Cai Yafei Y Wang G L GL

Cell stress & chaperones 20180120 4

The heat shock protein 90 (Hsp90) is a copious and ubiquitous molecular chaperone which plays an essential role in many cellular biological processes. The objective of this study was to identify single nucleotide polymorphisms (SNPs) in the Hsp90AA1 gene and to determine their association with heat stress traits in Chinese Holstein cattle breed. Direct sequencing was used to identify new SNPs. Luciferase reporter assay methods were used to assess g.- 87G > C and g.4172A > G loci in the promoter ...[more]

PMID: 29353403

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Project description:Heat shock proteins (Hsp) are known to protect cells from several stressors. Nucleotide changes in the flanking regions [5'- and 3'-untranslated region (UTR)] of Hsp gene might affect inducibility, degree of expression, or stability of Hsp70 mRNA. The present study aimed to investigate the association between inducible Hsp70.1 single nucleotide polymorphisms (SNPs) and heat shock (HS) response of peripheral blood mononuclear cells (PBMC) in dairy cows. Four hundred forty-six Italian Holstein cows were genotyped for four Hsp70.1 SNPs: g895 C/- and g1128 G/T in 5'-UTR, and g2154 G/A and g64 G/T in 3'-UTR. Genetic polymorphisms in 3'-UTR of bovine Hsp70.1 gene resulted monomorphic. Distribution of alleles of the nucleotide sequence polymorphism within the 5'-UTR of the bovine Hsp70.1 gene were 81.2% and 18.8% for C and -, respectively, and 77.8% and 22.2% for G and T, respectively. Among the 446 genotyped animals, a group of cows balanced for days in milk and parity was selected to be representative of the following genotypes: CC (n?=?8), C- (n?=?7), and -- (n?=?7) and GG (n?=?8), GT (n?=?11), and TT (n?=?3) in 5'-UTR. PBMC were isolated from blood samples and heated at 43°C in thermal bath for 1 h and then incubated at 39°C in atmosphere of 5% CO(2) for 1, 2, 4, 8, 16, and 24 h (recovery times). Cell viability was determined by XTT assay. Gene and protein expression of Hsp70.1 was determined by real-time reverse transcription-polymerase chain reaction and by ELISA assay, respectively. For the two SNPs detected, one allele was the most frequent (C, 66.8% and G, 56.8%). Genotypes -- and TG showed higher (P?<?0.05) viability compared with CC and GG, respectively. Genotypes C- and TT had intermediate viability. Gene expression of Hsp70.1 showed higher (P?<?0.001) levels in -- and TG genotype compared with their counterparts. Genotypes -- and TG showed the higher level of inducible Hsp70.1 protein in respect to C-, TT and CC, GG. In conclusion, exposure to HS differently affected cell viability and gene and protein expression of Hsp70.1 in the selected genotypes. These results indicate that the presence of SNPs (C/- and G/T) in the 5'-UTR region of inducible Hsp70.1 ameliorates HS response and tolerance to heat of bovine PBMC. These mutation sites may be useful as molecular genetic markers to assist selection for heat tolerance.

Dataset Information

Genetic polymorphism in Hsp90AA1 gene is associated with the thermotolerance in Chinese Holstein cows.

Publications

Genetic polymorphism in Hsp90AA1 gene is associated with the thermotolerance in Chinese Holstein cows.

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