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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제27권 제2호
- 발행연도
- 2017.1
- 수록면
- 357 - 364 (8page)
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In this study, we aimed to generate a series of versatile tagging plasmids that can be used in diverse molecular biological studies of the fungal pathogen Cryptococcus neoformans. We constructed 12 plasmids that can be used to tag a protein of interest with a GFP, mCherry, 4×FLAG, or 6×HA, along with nourseothricin-, neomycin-, or hygromycin-resistant selection markers. Using this tagging plasmid set, we explored the adenylyl cyclase complex (ACC), consisting of adenylyl cyclase (Cac1) and its associated protein Aca1, in the cAMP-signaling pathway, which is critical for the pathogenicity of C. neoformans. We found that Cac1-mCherry and Aca1-GFP were mainly colocalized as punctate forms in the cell membrane and nonnuclear cellular organelles. We also demonstrated that Cac1 and Aca1 interacted in vivo by coimmunoprecipitation, using Cac1-6×HA and Aca1-4×FLAG tagging strains. Bimolecular fluorescence complementation further confirmed the in vivo interaction of Cac1 and Aca1 in live cells. Finally, protein pull-down experiments using aca1Δ::ACA1-GFP and aca1Δ::ACA1- GFP cac1Δ strains and comparative mass spectrometry analysis identified Cac1 and a number of other novel ACC-interacting proteins. Thus, this versatile tagging plasmid system will facilitate diverse mechanistic studies in C. neoformans and further our understanding of its biology.
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참고문헌 (25)
참고문헌 신청
Alspaugh JA / 2002 / Adenylyl cyclase functions downstream of the Gα protein Gpa1 and controls mating and pathogenicity of Cryptococcus neoformans / Eukaryot. Cell 1 : 75 ~
Bahn YS / 2004 / Adenylyl cyclase-associated protein Aca1 regulates virulence and differentiation of Cryptococcus neoformans via the cyclic AMP-protein kinase A cascade / Eu
Bahn YS / 2013 / Stress signaling pathways for the pathogenicity of Cryptococcus / Eukaryot. Cell 12 : 1564 ~ 1577
Baker LG / 2012 / Multiple gene deletion in Cryptococcus neoformans using the cre-lox system / Methods Mol. Biol 845 : 85 ~ 98
Chang YC / 2012 / Role of actin-bundling protein Sac6 in
Bahn YS / 2004 / Adenylyl cyclase-associated protein Aca1 regulates virulence and differentiation of Cryptococcus neoformans via the cyclic AMP-protein kinase A cascade / Eu
Bahn YS / 2013 / Stress signaling pathways for the pathogenicity of Cryptococcus / Eukaryot. Cell 12 : 1564 ~ 1577
Baker LG / 2012 / Multiple gene deletion in Cryptococcus neoformans using the cre-lox system / Methods Mol. Biol 845 : 85 ~ 98
Chang YC / 2012 / Role of actin-bundling protein Sac6 in
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