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

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.

1. Alfirevic A., Mills T., Harrington P., Pinel T., Sherwood J., Jawaid A., Smith J.C., March R.E., Barratt B.J., Chadwick D.W., Kevin Park B., Pirmohamed M. Serious carbamazepine-induced hypersensitivity reactions associated with the HSP70 gene cluster. Pharmacogenet. Genomics, 2006, Vol. 16, no. 4, pp. 287-296.

2. Bogunia-Kubik K., Koscinska K., Suchnicki K., Lange A. HSP70-hom gene single nucleotide (+2763 G/A and +2437 C/T) polymorphisms in sarcoidosis. Int. J. Immunogenet., 2006, Vol. 33, no. 2, pp. 135-140.

3. Boyko A.A., Azhikina T.L., Streltsova M.A., Sapozhnikov A.M., Kovalenko E.I. HSP70 in human polymorphonuclear and mononuclear leukocytes: comparison of the protein content and transcriptional activity of HSPA genes. Cell Stress Chaperones, 2017, Vol. 22, no. 1, pp. 67-76.

4. Helmbrecht K., Zeise E., Rensing L. Chaperones in cell cycle regulation and mitogenic signal transduction: a review. Cell Proliferation, 2000, Vol. 33, no. 6, pp. 341-365.

5. Kampinga H.H., Hageman J., Vos M.J., Kubota H., Tanguay R.M., Bruford E.A., Cheetham M.E., Chen B., Hightower L.E. Guidelines for the nomenclature of the human heat shock proteins. Cell Stress Chaperones, 2009, Vol. 14, no. 1, pp. 105-111.

6. Kee C., Cheong K.Y., Pham K., Waterer G.W., Temple S.E. Genetic variation in heat shock protein 70 is associated with septic shock: Narrowing the association to a specific haplotype. Int. J. Immunogenet., 2008, Vol. 35, no. 6, pp. 465-473.

7. Martin A.M., Nolan D., Gaudieri S., Almeida C.A., Nolan R., James I., Carvalho F., Phillips E., Christiansen F.T., Purcell A.W., McCluskey J., Mallal S. Predisposition to abacavir hypersensitivity conferred by HLA-B*5701 and a haplotypic Hsp70-Hom variant. Proc. Natl. Acad. Sci. USA, 2004, Vol. 101, no. 12, pp. 4180-4185.

8. Maugeri N., Radhakrishnan J., Knight J.C. Genetic determinants of HSP70 gene expression following heat shock. Hum. Mol. Genet., 2010, Vol. 19, no. 24, pp. 4939-4947.

9. Shibata T., Arisawa T., Tahara T., Yoshioka D., Maruyama N., Fujita H., Kamiya Y., Nakamura M., Nagasaka M., Iwata M., Takahama K., Watanabe M., Hirata I., Nakano H. Protective role of genetic polymorphism of heat shock protein 70-2 for gastric cancer risk. Dig. Dis. Sci., 2009, Vol. 54, no. 1, pp. 70-74.

10. Spagnolo P., Sato H., Marshall S.E., Antoniou K.M., Ahmad T., Wells A.U., Ahad M.A., Lightman S., du Bois R.M., Welsh K.I. Association between heat shock protein 70/Hom genetic polymorphisms and uveitis in patients with sarcoidosis. Investig. Ophthalmol. Vis. Sci., 2007, Vol. 48, no. 7, pp. 3019-3025.

11. Temple S.E.L., Cheong K.Y., Ardlie K.G., Sayer D., Waterer G.W. The septic shock associated HSPA1B1267 polymorphism influences production of HSPA1A and HSPA1B. Intensive Care Med., 2004, Vol. 30, no. 9, pp. 1761-1767.

12. Ucisik-Akkaya E., Davis C.F., Gorodezky C., Alaez C., Dorak M.T. HLA complex-linked heat shock protein genes and childhood acute lymphoblastic leukemia susceptibility. Cell Stress Chaperones, 2010, Vol. 15, no. 5, pp. 475-485.