미생물학회지 (Korean Journal of Microbiology)

  • 제40권2호
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  • Pages.178-182
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  • 2004
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  • 0440-2413(pISSN)
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  • 2383-9902(eISSN)
한국미생물학회 (The Microbiological Society of Korea)
권호 표지

Aspergillus nidulans 영양균사에서 효소전구체형 ChsC 활성의 검출

Detection of Zymogenic ChsC Activity in Vegetative Hyphae of Aspergillus nidulans.

  • 박범찬 (충남대학교 생명과학부 미생물학전공) ;
  • 박윤희 (충남대학교 생명과학부 미생물학전) ;
  • 박희문 (충남대학교 생명과학부 미생물학전공)
  • 발행 : 2004.06.01
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초록

Saccharomyces cerevisiae의 class I 키틴생합성 효소인 Chsl의 활성측정법을 적용하여 Aspergillus nidulans의 영양균사에서 class I 키틴생합성 효소인 ChsC의 활성측정을 시도하였다. 그 결과, A. nidulans의 class I 키틴생합성효소도 효모류의 경우와 마찬가지로 트립신 처리에 의하여 활성화되는 효소전구체형태(zymogenic form)로 존재함을 알 수 있었다. 그리고 A. nidulans 야생형의 class I 키틴 생합성 효소활성은 트립신 처리에 의하여 6배 가량 증가되었다. 반면, chsC 유전자가 파괴된 돌연변이주는 트립신 처리에 의하여 효소활성이 증가되지 아니하였을 뿐만 아니라, 효소활성의 수준도 트립신을 처리하지 아니한 야생형의 class I 키틴생합성 효소활성과 거의 동일한 수준이었다. 따라서, 트립신을 처리하여 측정한A. nidulans 야생형의 class I 키틴생합성 효소활성 값에서 트립신을 처리하지 아니한 야생형의 class I 키틴생합성 효소활성을 제외한 값이 A. nidulans 야생형의 ChsC 효소활성임을 알았다. 이러한 조건을 토대로 영양균사 생장과정 동안 ChsC의 효소활성을 측정한 결과, chsC 유전자의 발현양상과 유사하게 액체배양상태의 영양균사가 무성분화능을 획득하는 시기로 알려진 시간대에 효소의 활성이 증가하였다. 이러한 결과는 이미 보고된 바와 같이 chs 유전자가 A. nidulans의 영양균사 생장에 관여함을 시사하고 있다.

In the vegetative hyphae of Aspergillus nidulans, a zymogenic form of the class I chitin synthase activity was successfully measured by the assay condition for Saccharomyces cerevisiae class I chitin synthase, Chsl. The class I chitin synthase activity of the A. nidulans chsC wild type strain was increased about six-fold by trypsin-pretreatment, but that of the chsC disruption strain revealed no increase. Interestingly enough, level of the class I chitin synthase activity of the chsC disruption strain was almost the same as that of the chsC wild type without trypsin-pretreatment. These results indicated that the A. nidulans ChsC activity could be measured by account-ing the class I chitin synthase activity without the trypsin-pretreatment as an internal control. Consistence to the expression pattern of the chsC revealed by northern blot analysis, the activity of ChsC was increased upon reaching the culture time for acquiring developmental competence. Our results shown here also supported the previous report suggesting the possible involvement of ChsC in vegetative hyphal growth of A. nidulans.

키워드

참고문헌

  1. Fugal Genet. Biol. v.20 The chsB gene of Aspergillus nidulansis necessary for normal hyphal growth and development Borgia,P.T.;N.Iartchouk;P.J.Riggle;K.R.Winter;Y.Koltin;C.E.Bulawa https://doi.org/10.1006/fgbi.1996.0035
  2. Cell v.46 The Saccharomyces cerevisiae structural gene for chitin synthase is not required for chitin synthesis in vivo Bulawa,C.E.;M.Slater;E.Cabib;J.Au Young;A.Sburlati;W.L.Jr.Adair;P.W.Robbins https://doi.org/10.1016/0092-8674(86)90738-5
  3. Anal. Biochem. v.219 The use of divalent cations and pH for the determination of specific yeast chitin synthetases Choi,W.J.;E.Cabib https://doi.org/10.1006/abio.1994.1278
  4. Biochem. Biophys. Res. Commun. v.236 A novel fungal gene encoding chitin synthase with a myosin motor-like domain Fujiwara,M.;H.Horiuchi;A.Ohta;M.Takagi https://doi.org/10.1006/bbrc.1997.6907
  5. J. Biochem. v.127 Evidence that the Aspergillus nidulans class I and class II chitin synthase genes, chsC and chsA, share critical roles in hyphal wall integrity and conidiophore development Fujiwara,M.;M.Ichinomiya;T.Motoyama;H.Horiuchi;A.Ohta;M.Takagi https://doi.org/10.1093/oxfordjournals.jbchem.a022616
  6. J. Bacteriol. v.181 Proliferation of intrahyphal hyphae caused by disruption of csmA, wich encodes a class V chitin synthase with a myosin motor-like domain in Aspergillus nidulans Horiuchi,H.;M.Fujiwara;S.Yamashita;A.Ohta;M.Takagi
  7. Micobiology v.148 Repression of chsβ expression reveals the functional importance of class IV chitin synthase gene chsD in hyphal growth and conidiation of Aspergillus nidulans Ichinomiya,M.;T.Motoyama;M.Fujiwara;M.Takagi;H.Horiuchi;A.Ohta
  8. Eur. J. Biochem. v.269 Involvement of GFAL,wich encodes glutamine-fructose-6-phosphate amidotransferase, in the activation of the chitin synthesis pathway in response to cell-wall defects in Saccharomyces cerevisiae Lagorce,A.;V.Le Berre Anton;B.Aguilar Uscanga;H.Martin Yken;A.Dagkessamanskaia;J.Francois https://doi.org/10.1046/j.1432-1327.2002.02814.x
  9. Fungal Genet. Biol. v.41 Differential expreesion of the chitin synthase genes of Aspergillus nidulans. chsA, chsB, and chsC, in response to developmental status and environmental factors Lee,J.L.;J.H.Choi;B.C.Park;Y.H.Park;M.Y.Lee;H.M.Park;P.J.Maeng https://doi.org/10.1016/j.fgb.2004.01.009
  10. Biosci. Biotechnol. Biochem. v.58 Isolation of a chitin synthase gene(chsC)of Aspergillus nidulans Motoyama,T.;M.Kojima;H.Horiuchi;A.Ohta;M.Tagagi https://doi.org/10.1271/bbb.58.2254
  11. J. Biol. Chem. v.262 Two chitin synthase in Saccharomyces cerevisiae Orlean,P.
  12. J. Microbiol. v.39 Cell cycle-dependent expression of chitin synthase genes in Aspergillu nidulans Park,B.C.;P.J.Maeng;H.M.Pakr
  13. FEMS Microbiol. Lett. v.220 Activation of chsC transcription by AbaA during asexual development of Aspergillus nidulans Park,B.C.;Y.H.Park;H.M.Park https://doi.org/10.1016/S0378-1097(03)00120-4
  14. Ph.D.Thesis. Chungnam National University Park,B.C.
  15. Curr. Genet. v.41 The genetic complexity of chitin synthesis in fungi Roncero,C. https://doi.org/10.1007/s00294-002-0318-7
  16. J. Biol. Chem. v.261 Chitin synthetase2, a presumptive participant in septum formation in Saccharomyces cerevisiae Sburlatia,A.;E.Cabib
  17. J. Cell Biol. v.114 The function of chitin synthases 2 and 3 in Saccharomyces cerevisiae cell cycle Shaw,J.A.;P.C.Mol;B.Bower;S.J.Silverman;M.H.Valdivieso;A.Duran;E.Cabib https://doi.org/10.1083/jcb.114.1.111
  18. Fungal Genet. Biol. v.20 The chsD and chsE genes of Aspergillus nidulans and their roles in chitin synthesis Specht,C.A.;Y.Liu;P.W.Robbins;C.E.Bulawa;N.Iartchouk;K.R.Winter;P.J.Riggle;J.C.Rhodes;C.L.Dodge;D.W.Culp;P.T.Borgia https://doi.org/10.1006/fgbi.1996.0030
  19. J. Bacteriol. v.182 Chitin synthesis in gasl mutant of Saccharomyces cerevisiae Valdivieso,M.H.;L.Ferrario;M.Vai;A.Duran;L.Popolo https://doi.org/10.1128/JB.182.17.4752-4757.2000