SURFACE PLASMON RESONANCE STUDY OF THE INTERACTION BETWEEN THE HUMAN TATA-BOX BINDING PROTEIN AND THE TATA ELEMENT OF THE NOS2A GENE PROMOTER

Interaction ofthe TATA-binding protein (ТВР) with the TATA box and formation ofthe TBP/TATA complex is the first step in the transcription complex assembly and the central event in transcription regulation. It is known that some single nucleotide polymorphisms (SNPs) in the human TATA box and its flanks are associated with elevated risk ofvarious disorders (Savinkova etal., 2009). Little is known about changes in the TBP/TATA interaction leading to the elevated risk ofany disease. Thus, we have studied the influence ofSNP (–21 t > c), adjoining to the TATA box ofthe NOS2A promoter, on the interaction by surface plasmon resonance (SPR) on biosensor ProteOn XPR36 (BioRad). The sensograms obtained in the experiment indicate that the constant for the interaction with TATA bearing SNP –21 t > c differ little from the constant with the wild-type box and the dissociation constants differ approximately twofold. Experiments utilizing the SPR method and classical EMSA have shown that ТВР immobilization on the microchip dextran surface does not alter its affinity to TATA-containing oligonucleotides.

1. Драчкова И.А., Аршинова Т.В., Пономаренко П.М. и др. Влияние полиморфизмов ТАТА-бокса промотора гена β-глобина человека, ассоциированных с β-талассемией, на взаимодействие ТАТА-связывающего белка // Информ. вестник ВОГиС. 2010. Т. 14. № 4. С. 698–705.

2. Савинкова Л.К., Пономаренко М.П., Пономаренко П.М. и др. Полиморфизмы ТАТА-боксов промоторов генов человека и ассоциированные с ними наследственные патологии // Биохимия. 2009. Т. 74. № 4. С. 149–163.

3. Соколенко А.А., Сандомирский И.И., Савинкова Л.К. Взаимодействие дрожжевого ТАТА-связывающего белка с корткими участками промоторов // Молекуляр. биология. 1996. Т. 30. № 2. С. 279–285.

4. Abbas A., Linman M.J., Cheng C. New trends in instrumental design for surface plasmon resonance-based biosensors // Biosens. Bioelectron. 2011. V. 26. No. 5. P. 1815–1824.

5. Abdiche Y.N., Lindquist K.C., Pinkerton A. et al. Expanding the proteOn XPR36 biosensor into a 36-ligand array expedites protein interaction analysis // Analyt. Biochem. 2011. V. 411. P. 139–151.

6. Barcellos L.F., Ramsay P.P, Caillier S.J. et alal. Genetic variation in nitric oxide synthase 2A (NOS2A) and risk for multiple sclerosis // Genes Immun. 2008. V. 9. P. 493–500.

7. Bronner V., Denkberg G., Peled M. et al. Therapeutic antibodies: Discovery and development using the ProteOn XPR36 biosensor interaction array system // Analyt. Biochem. 2010. V. 406. P. 147–156.

8. Burgner D., Rockett K., Ackerman H. et alal. Haplotypic relationship between SNP and microsatellite markers at the NOS2A locus in two // Genes Immun. 2003. V. 4. P. 506–514.

9. Juven-Gershon T., Kadonaga J.T. Regulation of gene expression via the core promoter and the basal transcriptional machiner // Dev. Biol. 2010. V. 339. No. 2. P. 225–229.

10. Kim Y., Geiger J.H., Hahn S., Sigler P.B. Crystal structure of a yeast TBP/TATA-box complex // Nature. 1993. V. 365. P. 512–520.

11. Nahshol O., Bronner V., Notcovich A. et al. Parallel kinetic analysis and affinity determination of hundreds of monoclonal antibodies using the ProteOn XPR36 // Anal. Biochem. 2008. V. 383. No. 1. P. 52–60.

12. Navratilova I., Dioszegi M., Myszka D.G. Analyzing ligand and small molecule binding activity of solubilized GPCRs using biosensor technology // Analyt. Biochem. 2006. V. 355. P. 132–139.

13. Rich R.L., Myszka D.G. Grading the commercial optical biosensor literature-Class of 2008: ‘The Mighty Binders // Mol. Recognit. 2010. V. 23. P. 1–64.

14. Strahs D., Barash D., Qian., Schlick T. Sequence-dependent solution structure and motions of 13 TATA/TBP (TATA-box binding protein) complexes // Biopolymers. 2003. V. 69. P. 216–243.