Analysis of the association of the polymorphism of the CLIC1, MSH5, C6orf26, C6orf25 genes with the expression level of the HSPA1B gene

Abstract

Heat shock proteins (HSP, heat shock proteins) form one of the cellular molecular systems with chaperone activity, aimed at stabilizing the structure of intracellular proteins, ensuring the resistance of cells to stress, renaturation of incorrectly folded and elimination of denatured intracellular proteins.Our task was to look for an association between the presence of single nucleotide polymorphisms in selected regions of the genome and the basal level of transcriptional activity of the HSPA group genes, namely HSPA1A/B, HSPA1A, HSPA1B, HSPA6 and HSPA8 in mononuclear leukocytes (PBMC), analyzed in our experiments volunteers from the population of the middle part of Russia in order to analyze the universality of the biological effects of these SNPs.The study was performed on DNA and RNA isolated from peripheral blood lymphocytes of 16 donors. Genotyping was performed by the polymerase chain reaction (PCR) followed by sequencing of the PCR product. To assess the level of gene expression of the HSPA group, cDNA was synthesized on an RNA template isolated from PBMC cell fraction samples, followed by real-time polymerase chain reaction.The following types of polymorphisms were genotyped: rs400547 (A/G), rs1150793 (G/A), rs707936 (A/G), rs707915 (A/T), rs376510 (T/C) located in the CLIC1, MSH5, C6orf26, MSH5, C6orf25genes, respectively. We determined the basal level of transcription of genes of constitutively expressed and inducible proteins of the HSP70 family and searched for the association of their expression with polymorphisms.It was found that in PBMC cells, DNA that has the following genotype: AG/AA (SNP rs400547), AG/GG (rs1150793), AG (rs707936), TArs707915, TC (rs376510), in the regions we studied, is associated with a decrease in the transcription of the HSPA1B gene (p = 0.02) compared with homozygotes: GG (SNP rs400547), AA (rs1150793), GG (rs707936), TT (rs707915), CC (rs376510).Associations of these polymorphisms with gene expression of HSPA1A, HSPA6 and HSPA8 have not been identified.The CLIC1, MSH5, C6orf26, C6orf25 genes, in which the polymorphisms studied by us are present, are located in the same locus near the HSPA1B gene on the 6th chromosome. We found a decrease in HSPA1B gene expression in the presence of single nucleotide polymorphisms in nearby genes may indicate spatial interactions of this locus and the HSPA1B gene locus, and that a change in the genotype CLIC1, MSH5, C6orf26, C6orf25 may entail a change in the expression of closely arranged genes which are functionally significant for the cell.

Keywords

• snps
• heat shock proteins
• hspa1b gene