Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Identification and Functional Analysis of Proteins Interacting with Streptomyces coelicolor RNase ES

Streptomyces coelicolor 리보핵산내부분해효소 RNase ES의 결합단백질 규명 및 기능분석

  • 김종명 (중앙대학교 자연과학대학 생명과학과) ;
  • 송우석 (중앙대학교 자연과학대학 생명과학과) ;
  • 김현리 (중앙대학교 자연과학대학 생명과학과) ;
  • 고하영 (중앙대학교 자연과학대학 생명과학과) ;
  • 이강석 (중앙대학교 자연과학대학 생명과학과)
  • Published : 2007.03.31
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Abstract

Using co-immunoprecipitation, we identified proteins interacting with Streptomyces coelicolor RNase ES, an ortholog of Escherichia coli RNase E that plays a major role in RNA decay and processing. Polyphosphate kinase and a homolog of exoribonuclease polynucleotide phosphorylase, guanosine pentaphosphate synthetase I that use inorganic phophate were co-precipitated with RNase E, indicating a possibility of S. coelicolor RNase ES to form a multiprotein complex called degradosome, which has been shown to be formed by RNase E in E. coli. Polynucleotide phophorylase proteins from these two phylogenetically distantly related bacteria species showed similar RNA cleavage action in vitro. These results imply the ability of RNase ES to form a multiprotein complex that has structurally and functionally similar to that of E. coli degradosome.

Escherichia coli에서 RNA 분해와 가공에 있어서 중심적인 역할을 하는 RNase E의 동족체 단백질인 Streptomyces coelicolor RNase ES의 결합 단백질을 항체침전을 이용하여 탐색하였다. 무기인산을 이용하는 polyphosphate kinase와 리보핵산외부분해효소인 polynucleotide phophorylase의 동족체인 GPSI가 RNase ES와 함께 침전되는 것을 확인하였으며, 이는S. coelicolor에도 E. coli RNase E를 매개로 구성되는 다단백질복합체인 degradosome이 RNase ES에 의해 형성될 수 있음을 암시한다. 계통적으로 멀리 떨어진 이 두 세균에서 정제된 polynucleotide phosphorylase 동족체는 시험관에서의 RNA 분해 특성이 서로 유사함을 보였다. 이러한 실험 결과는 RNase ES가 E. coli degradosome과 유사한 기능과 구조를 가진 단백질 복합체를 형성함을 시사한다.

Keywords

References

  1. Apirion, D. and A.B. Lassar. 1978. A conditional lethal mutant of Escherichia coli which affects the processing of ribosomal RNA. J. Biol. Chem. 253, 1738-1742
  2. Carpousis, A.J., G. Van Houwe, C. Ehretsmann, and H.M. Krisch. 1994. Copurification of E. coli RNAse E and PNPase: evidence for a specific association between two enzymes important in RNA processing and degradation. Cell 76, 889-900 https://doi.org/10.1016/0092-8674(94)90363-8
  3. Condon, C., J. Rourera, D. Brechemier-Baey, and H. Putzer. 2002. Ribonuclease M5 has few, if any, mRNA substrates in Bacillus subtilis. J. Bacteriol. 184, 2845-2849 https://doi.org/10.1128/JB.184.10.2845-2849.2002
  4. Gao, J., K. Lee, M. Zhao, J. Qiu, X. Zhan, A. Saxena, C.J. Moore, S.N. Cohen, and G. Georgiou. 2006. Differential modulation of E. coli mRNA abundance by inhibitory proteins that alter the composition of the degradosome. Mol. Microbiol. 61, 394-406 https://doi.org/10.1111/j.1365-2958.2006.05246.x
  5. Gurevitz, M. and D. Apirion. 1983. Interplay among processing and degradative enzymes and a precursor ribonucleic acid in the selective maturation and maintenance of ribonucleic acid molecules. Biochemistry 22, 4000-4005 https://doi.org/10.1021/bi00286a002
  6. Jger, S., O. Fuhrmann, C. Heck, M. Hebermehl, E. Schiltz, R. Rauhut, and G. Klug 2001. An mRNA degrading complex in Rhodobacter capsulatus. Nucleic Acids Res. 29, 4581-4588 https://doi.org/10.1093/nar/29.22.4581
  7. Jones, G.H. and M.J. Bibb. 1996. Guanosine pentaphosphate synthetase from Streptomyces antibioticus is also a polynucleotide phosphorylase. J. Bacteriol. 178, 4281-8 https://doi.org/10.1128/jb.178.14.4281-4288.1996
  8. Kim, S., H. Kim, I. Park, and Y. Lee. 1996. Mutational analysis of RNA structures and sequences postulated to affect 3' processing of M1 RNA, the RNA component of Escherichia coli RNase P. J. Biol. Chem. 271, 19330-19337 https://doi.org/10.1074/jbc.271.32.19330
  9. Lee, K. and S.N. Cohen. 2001. Effect of 3'terminus modifications on mRNA functional decay during in vitro protein synthesis. J. Biol. Chem. 276, 23268-23274 https://doi.org/10.1074/jbc.M102408200
  10. Lee, K, J.A. Bernstein, and S.N. Cohen. 2002. RNase G complementation of me null mutation identifies functional interrelationships with RNase E in Escherichia coli. Mol. Microbiol. 43, 1445-1456 https://doi.org/10.1046/j.1365-2958.2002.02848.x
  11. Lee, K. and S.N. Cohen. 2003. A Streptomyces coelicolor functional orthologue of Escherichia coli RNase E shows shuffling of catalytic and PNPase-binding domains. Mol. Microbiol. 48, 349-360 https://doi.org/10.1046/j.1365-2958.2003.03435.x
  12. Lee, K., X. Zhan, J. Gao, J. Qui, Y. Feng, R. Meganathan, S.N. Cohen, and G. Giorgiou. 2003. RraA a protein inhibitor of RNase E activity that globally modulates RNA abundance in E. coli. Cell. 114, 623-634
  13. Leroy, A., N.F. Vanzo, S. Sousa, M. Dreyfus, and A.J. Carpousis. 2002. Function in Escherichia coli of the non-catalytic part of RNase E: role in the degradation of ribosome-free mRNA Mol. Microbiol. 45, 1231-1243 https://doi.org/10.1046/j.1365-2958.2002.03104.x
  14. Li, Z., S. Pandit, and M.P. Deutscher. 1999. RNase G(CafA protein) and RNase E are both required for the 5' maturation of 16S ribosomal RNA. EMBO J. 18, 2878-2885 https://doi.org/10.1093/emboj/18.10.2878
  15. Liou, G.G., W.N. Jane, S.N. Cohen, N.S. Lin, and S. Lin-Chao. 2001. RNA degradosome exist in vivo in Escherichia coli as multicomponent complexes associated with the cytoplasmic membrane via the N-terrnianal region of ribonuclease E. Proc. Natl. Acad. Sci. USA. 98, 63-68
  16. McDowall, K.J. and S.N. Cohen. 1996. The N-terminal domain of the me gene product has RNase E activity and is non-overlapping with the arginine-rich RNA-binding site. J. Mol. Biol. 255, 349-355 https://doi.org/10.1006/jmbi.1996.0027
  17. Miczak, A., V.R. Kaberdin, C.L. Wei, and S. Lin-Chao. 1996. Proteins associated with RNase E in a multicomponent ribonucleolytic complex. Proc. Natl. Acad. Sci. USA. 90, 3865-3869
  18. Morita, T., H. Kawamoto, T. Mizota, T. Inada, and H. Aiba. 2004. Enolase in the RNA degradosome plays a crucial role in the rapid decay of glucose transporter mRNA in the response to phosphosugar stress in Escherichia coli. Mol. Microbiol. 54, 1063-1075 https://doi.org/10.1111/j.1365-2958.2004.04329.x
  19. Py, B., C.F. Higgins, H.M. Krisch, and A.J. Carpousis. 1996. A DEAD-box RNA helicase in the Escherichia coli RNA degradosome. Nature 381, 169-172 https://doi.org/10.1038/381169a0
  20. Vanzo, N.F., Y.S. Li, B. Py, E. Blem, C.F. Higgins, L.C. Raynal, H.M. Krisch, and A,J. Carpousis. 1998. Ribonuclease E organizes the protein interactions in the Escherichia coli RNA degradosome. Genes Dev. 12, 2770-2781 https://doi.org/10.1101/gad.12.17.2770