• 제목/요약/키워드: Yeast Two-hybrid

검색결과 231건 처리시간 0.032초

분열효모에서 hnRNP-유사 단백질인 THO4가 생장 및 mRNA 방출에 미치는 영향 (Effect of hnRNP-like protein THO4 on growth and mRNA export in fission yeast)

  • 박진희;이소정;윤진호
    • 미생물학회지
    • /
    • 제54권2호
    • /
    • pp.91-97
    • /
    • 2018
  • 진화적으로 보존된 TREX 복합체의 구성요소인 Yra1/ALY는 hnRNP-유사 단백질의 REF (RNA와 방출인자 결합단백질) 계열에 속하는 단백질로서 전사, 핵 RNA의 안정성, mRNA의 핵에서 방출 등 여러 과정에 관여하는 것으로 알려져 있다. 분열효모 Schizosaccharomyces pombe의 게놈에는 2개의 REF 단백질을 암호화하고 있다. 이미 mRNA 방출인자로 알려진 Mlo3 이외에 THO 복합체의 구성인자로 예측되는 Tho4이 존재한다. 본 연구에서 tho4 (SPBC106.12c) 유전자의 결실이 생장과 mRNA의 방출에 영향을 주지 않는다는 것을 알았다. 그러나 tho4 유전자를 과발현시키면 생장이 늦어지고 $poly(A)^+$ RNA가 핵 안에 약간 축적되었다. ${\Delta}tho4$ ${\Delta}mlo3$${\Delta}tho4$ ${\Delta}mex67$ 이 중 돌연변이는 어느 것도 추가적인 생장 결함을 보이지 않았다. 또한 Yeast two-hybrid와 공동침전(Co-immunoprecipitation) 분석에서 Tho4는 THO/TREX 복합체의 다른 구성인자들과 어떠한 상호작용도 보이지 않았다. 이와 같은 관찰결과들은 S. pombe의 Tho4 단백질이 기대와는 다르게THO/TREX 복합체의 구성인자가 아니며, mRNA 방출에도 직접 관여하지 않음을 의미한다.

Isolation and Characterization of a cDNA Encoding Two Novel Heat-shock Factor OsHSF6 and OsHSF12 in Oryza Sativa L.

  • Liu, Jin-Ge;Yao, Quan-Hong;Zhang, Zhen;Peng, Ri-He;Xiong, Ai-Sheng;Xu, Fang;Zhu, Hong
    • BMB Reports
    • /
    • 제38권5호
    • /
    • pp.602-608
    • /
    • 2005
  • As a crucial transcription factor family, heat-shock factors were mainly analyzed and characterized in tomato and Arabidopsis. In this study, we isolated two putative heat shock factors OsHSF6 and OsHSF12 that interact specifically with heat-shock element (HSE) from Oryza sativa L by yeast one-hybrid method. The full-length cDNA of OsHSF6 and OsHSF12 have 1074bp and 920bp open reading frame (ORF), respectively. Analysis of the deduced amino acid sequences revealed that OsHSF6 was a class A heat shock factor (HSF) with all the conserved sequence elements characteristic of heat stress transcription factor, while OsHSF12 was a class B HSF with C-terminal domain (CTD) lacking of AHA motif. Bioinformatic analysis showed that the sequences and structures of two HSFs' DNA binding domain (DBD) had a high similarity with LpHSF24. The results of RT-PCR indicated OsHSF6 gene was expressed immediately after rice plants exposure to heat stress, and the transcription of OsHSF6 gene accumulated primarily in immature seeds, roots and leaves. However, we did not find the transcription of OsHSF12 gene in different organs and growth periods. Our results implied that OsHSF6 might be function as a HSF regulating early expression of stress genes in response to heat shock, and OsHSF12 might be act as a synergistic factor to regulate the expression of down-stream genes.

RPK118, a PX Domain-containing Protein, Interacts with Peroxiredoxin-3 through Pseudo-Kinase Domains

  • Liu, Lungling;Yang, Chenyi;Yuan, Jian;Chen, Xiujuan;Xu, Jianing;Wei, Youheng;Yang, Jingchun;Lin, Gang;Yu, Long
    • Molecules and Cells
    • /
    • 제19권1호
    • /
    • pp.39-45
    • /
    • 2005
  • RPK118 is a sphingosine kinase-1-binding protein that has been implicated in sphingosine 1 phosphate-mediated signaling. It contains a PX (phox homology) domain and two pseudo-kinase domains, and co-localizes with sphingosine kinase-1 on early endosomes. In this study we identified a novel RPK118-binding protein, PRDX3 (peroxiredoxin-3), by yeast two-hybrid screening. The interaction between these proteins was confirmed by pull-down assays and co-immunoprecipitation experiments. Deletion studies showed that RPK118 interacted with PRDX3 through its pseudokinase domains, and with early endosomes through its PX domain. Double immunofluorescence experiments demonstrated that PRDX3 co-localized with RPK118 on early endosomes in COS7 cells. PRDX3 is a member of the antioxidant family of proteins synthesized in the cytoplasm and functioning in mitochondria. Our findings indicate that RPK118 is a PRDX3-binding protein that may be involved in transporting PRDX3 from the cytoplasm to its mitochondrial site of function or to other membrane structures via endosome trafficking.

Glutamic Acid Rich Helix II Domain of the HIV-1 Vpu has Transactivation Potential in Yeast

  • Hong, Seung-Keun;Bae, Yong-Soo;Kim, Jung-Woo
    • BMB Reports
    • /
    • 제32권4호
    • /
    • pp.405-408
    • /
    • 1999
  • The transactivation potential of HIV-1 Vpu was identified from the yeast two-hybrid screening process. The helix II domain of HIV-1 Vpu protein and mutant Vpu protein lacking the transmembrane domain exhibited transactivation of the LacZ and Leu2 reporter genes carrying LexA upstream activating sequences, but full-length HIV-1 Vpu and the helix I domain of HIV-1 Vpu did not. The helix II domain of HIV-1 Vpu consists of a number of acidic amino acids, and is especially rich in glutamic acid, a characteristic of many transcription factors. This result suggests that protein-protein interaction may occur through the acidic helix II domain of HIV-1 Vpu.

  • PDF

Role of CaBud6p in the Polarized Growth of Candida albicans

  • Song Yun-Kyoung;Kim Jeong-Yoon
    • Journal of Microbiology
    • /
    • 제44권3호
    • /
    • pp.311-319
    • /
    • 2006
  • Bud6p is a component of a polarisome that controls cell polarity in Saccharomyces cerevisiae. In this study, we investigated the role of the Candide albicans Bud6 protein (CaBud6p) in cell polarity and hyphal development. CaBud6p, which consists of 703 amino acids, had 37% amino-acid sequence identity with the Bud6 protein of S. cerevisiae. The homozygous knock-out of CaBUD6 resulted in several abnormal phenotypes, such as a round and enlarged cells, widened bud necks, and a random budding pattern. In hypha-inducing media, the mutant cells had markedly swollen tips and a reduced ability to switch from yeast to hypha. In addition, a yeast two-Hybrid analysis showed a physical interaction between CaBud6p and CaAct1p, which suggests that CaBud6p may be involved in actin cable organization, like Bud6p in S. cerevisiae. Taken together, these results indicate that CaBud6 plays an important role in the polarized growth of C. albicans.

Kinesin superfamily KIF21A와 직접 결합하는 Pcp-2의 규명 (Pcp-2 Interacts Directly with Kinesin Superfamily KIF21A Protein)

  • 박혜영;김상진;예성수;장원희;이상경;박영홍;정용욱;문일수;김무성;석대현
    • 생명과학회지
    • /
    • 제18권8호
    • /
    • pp.1059-1065
    • /
    • 2008
  • KIF21A는 kinesin superfamily에 속하는 분자 motor로서 미세소관을 따라서 분비소포를 운반한다. 최근의 연구결과 KIF21A 유전자 일부의 missense 돌연변이에 의하여 congenital fibrosis of the extraocular muscles (CFEOM) 1의 유발됨이 밝혀졌다. CFEOM1은 KIF21A의 돌연변이로 인하여 분화 발생과정에 occulo-motor신경과 neuromuscular junction 형성에 필요한 단백질을 이동시키지 못함으로써 유발된다. 본 연구에서는 효모 two-hybrid system을 사용하여 KIF21A의 WD-40 repeat domain과 결합하는 분자량이 작은 Purkinje cell protein-1 (Pcp-2), 또는 L7으로도 알려진 단백질을 분리하였다. Pcp-2는 효모 two-hybrid assay에서 KIF21A와 KIF21B의 WD-40 영역과는 결합하지만 다른 종류의 KIFs와는 결합하지 않았다. 또한 단백질간의 특이적 결합을 pull-down assay로 확인하였으며, 생쥐의 뇌 파쇄액에 Pcp-2 항체로 면역침강을 행하여 KIF21A를 확인한 결과 Pcp-2와 같이 침강하였다. 이러한 결과들은 KIF21A는 Pcp-2와 결합하며, 또한 Pcp-2는 KIF21A의 adaptor 단백질로서 세포 내 KIF21A의 수송에서 매개 단백질로 작용함을 시사한다.

내분비계 장애물질의 개요와 검색법의 고찰 (Overall Review on endocrine disruptors)

  • 류재천
    • 농약과학회지
    • /
    • 제6권3호
    • /
    • pp.135-156
    • /
    • 2002
  • 내분비계 장애물질이 환경호르몬이란 용어로 우리들의 실생활에 사용되는 용기 등을 통해 여러 가지 영향을 미치고 있다. 수많은 화학물질들 중에서 환경호르몬은 어떠한 종류의 화학물질이며, 언제 알려지게 되었고, 어떻게 분류되었으며, 이러한 환경호르몬성을 갖는 물질들의 환경호르몬성은 어떠한 연구기법에 의해 검색되는지와 현재까지의 사용실태와 문제점 및 국내외 관련 동향 등을 고찰하여 보았다.