생명과학회지 (Journal of Life Science)

  • 제16권7호
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  • Pages.1225-1228
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  • 2006
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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DNA 벤딩(휨) 없이 돌연변이 cAMP 수용체 단백질의 결합

Mutant cAMP Receptor Protein Binds to DNA without DNA Bending

  • 발행 : 2006.12.01
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⟨ 이전 논문 다음 논문 ⟩

초록

cAMP와 복합체를 형성한 cAMP 수용성 단백질은 DNA와 결합하여 ${\sim}90$도 정도의 예리한 DNA bending을 유도한다. 그러나 이전의 논문[5]에 의하면 돌연변이 CRP:cGMP 복합체는 돌연변이 CRP:cAMP 복합체보다 아크릴아미드 겔에서 상대적으로 빠른 이동속도를 보였다. CRP와 cyclic nucleotide 존재하에서 DNA의 구조 변화를 알아보기 위하여 6가지 준비된 DNA조각들을 사용하여 몰 고리화 인자(molar cyclization factor)[13]를 측정하였다. 이들 자료를 사용하여 nonlinear regression analysis를 통하여 cGMP 존재하에서 돌연변이 CRP는 DNA bending을 형성하지 않으나 CAMP 존재하에서 나선 꼬임과 같은 DNA 구조 변화없이 DNA bending을 형성한다.

Cyclic AMP receptor protein (CRP) complexed with cAMP binds to DNA and induces sharp DNA bending around ${\sim}90$ degree. Previous publication (5), however, reported that mutant CRP:cGMP complex showed high migration rate relative to mutant CRP:cAMP complex on native polyacrylamide gel. To confirm DNA structural change in the presence of CRP and cyclic nucleotide, molar cyclization factor $(j_M)$ [13] was measured with 6 constructed DNA fragments. Nonlinear regression analysis of $j_M$ data indicated that mutant CRP did not induce DNA bending in the presence of cGMP but bent DNA in the presence of cAMP without any helical twist change in DNA.

키워드

참고문헌

  1. deCrombrugghe, B., S. Busby, and H. Buc. 1984. Cyclic AMP receptor protein: role in transcription activation. Science 224, 831-838 https://doi.org/10.1126/science.6372090
  2. Dripps, D. and R. M. Wartell. 1987. DNA bending induced by the catabolite activator protein allows ring formation of 144 bp DNA. J. Biomol. Struct. Dyn. 5, 001-012 https://doi.org/10.1080/07391102.1987.10506370
  3. Gang, J. 1994. Cooperative binding of E. coli RNA polymerase and a mutant cAMP receptor protein to the lactose promoter. Korean Biochem. J. 27, 350-352
  4. Gang, J. 2004. Measurement of DNA helical change for the binding of cyclic AMP receptor protein to lac DNA. Biochem. Biophys. Res. Commun. 322, 993-997 https://doi.org/10.1016/j.bbrc.2004.08.011
  5. Gang, J. and J. G. Harman. 1997. The characteristic binding of mutant cyclic AMP receptor protein (CRP*) to lactose promoter in Escherichia coli. Mol. Cells 4, 167-170
  6. Harman, J. G. 2001. Allosteric regulation of the cAMP receptor protein. Biochim. Biophys. Acta. 1547, 1-17 https://doi.org/10.1016/S0167-4838(01)00187-X
  7. Harman, J. G., K. Mckenney, and A. Petrokofsky. 1986. Structure-function analysis of three cAMP-independent forms of the cAMP receptor protein. J. Biol Chem. 261, 16332-16339
  8. Heyduk, T. and J. C. Lee. 1992. Solution studies on the structure of bent DNA in the cAMP receptor protein:lac DNA complex. Biochemistry 31, 5165-5171 https://doi.org/10.1021/bi00137a011
  9. Kolb, A., S. Busby, H. Buc, S. Garges, and S. Adhya. 1993. Transcriptional regulation by cAMP and its receptor protein. Ann. Rev. Biochem. 62, 749-795 https://doi.org/10.1146/annurev.bi.62.070193.003533
  10. Kotlarz, D., A. Fritsch, and H. Buc. 1986. Variations of the intramolecular ligation rates allow the detection of protein-induced bends in DNA. EMBO J. 5, 799-803
  11. Pyles, E. A. and J. C. Lee. 1998. Escherichia coli cAMP receptor protein-DNA complexes. Biochemistry 37, 5201-5210 https://doi.org/10.1021/bi972451a
  12. Reznikoff, W. S. 1992. Catabolite gene activator protein activation of lac transcription. J. Bacterial. 174, 655-658 https://doi.org/10.1128/jb.174.3.655-658.1992
  13. Taylor, W. H. and P. J. Hagerman. 1990. Application of the method of phage T4 DNA ligase-catalyzed ring-closure to the study of DNA structure. J. Mol. Biol. 212, 363-376 https://doi.org/10.1016/0022-2836(90)90131-5