Functional and physical interactions between components of the Prp19p-associated complex.
The Prp19p-associated complex is essential for the yeast pre-mRNA splicing reaction. The complex consists of at least eight protein components, but is not tightly associated with spliceosomal snRNAs. By a combination of genetic and biochemical methods we previously identified four components of this complex, Ntc25p, Ntc85p, Ntc30p and Ntc20p, all of them being novel splicing factors. We have now identified three other components of the complex, Ntc90p, Ntc77p and Ntc31p. These three proteins were also associated with the spliceosome during the splicing reaction in the same manner as Prp19p, concurrently with or immediately after dissociation of U4 snRNA. Two-hybrid analysis revealed that none of these proteins interacted with Prp19p or Ntc25p, but all interacted with Ntc85p. An interaction network between the identified components of the Prp19p-associated complex is demonstrated. Biochemical analysis revealed that Ntc90p, Ntc31p, Ntc30p and Ntc20p form a subcomplex, which, through interacting with Ntc85p and Ntc77p, can associate with Prp19p and Ntc25p to form the Prp19p-associated complex. Genetic analysis suggests that Ntc31p, Ntc30p and Ntc20p may play roles in modulating the function of Ntc90p.
Chen
Chun-Hong
CH
Institute of Molecular Biology, Academia Sinica, Nankang, Taiwan, Republic of China.
Yu
Wan-Chin
WC
Tsao
Twee Y
TY
Wang
Lian-Yung
LY
Chen
Hau-Ren
HR
Lin
Jui-Yen
JY
Tsai
Wei-Yü
WY
Cheng
Soo-Chen
SC
D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D002455:Cell Division; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D046911:Macromolecular Substances; D008954:Models, Biological; D009154:Mutation; D011233:Precipitin Tests; D012326:RNA Splicing; D000072260:RNA Splicing Factors; D012331:RNA, Fungal / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D017381:Spliceosomes / Q000378:metabolism; D020798:Two-Hybrid System Techniques
C419763:CLF1 protein, S cerevisiae; D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D046911:Macromolecular Substances; C079877:PRP19 protein, S cerevisiae; D000072260:RNA Splicing Factors; D012331:RNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; C419764:Syf1 protein, S cerevisiae