Nucleic Acids Research, 2002, Vol. 30, No. 4 1029-1037
© 2002 Oxford University Press
Functional and physical interactions between components of the Prp19p-associated complex
1Institute of Molecular Biology, Academia Sinica, Nankang, Taiwan, Republic of China and 2Institute of Microbiology and Immunology, National Yang-Ming University, Shih-Pai, Taiwan, Republic of China
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.
* To whom correspondence should be addressed at: Institute of Molecular Biology, Academia Sinica, 128 Academia Road, Section 2, Nankang, Taiwan, Republic of China. Tel: +886 2 27899200; Fax: +886 2 27883296; Email: mbscc{at}ccvax.sinica.edu.tw Present addresses:Wan-Chin Yu, Process Development Division, Development Center for Biotechnology, Taipei, Taiwan, Republic of ChinaHau-Ren Chen, Institute of Molecular Virology, Howard Hughes Medical Institute, University of Wisconsin, Madison, WI 53706, USAWei-Yü Tsai, School of Medical Technology, Chung Shan Medical and Dental College, Taichung, Taiwan, Republic of China
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