TY - JOUR
T1 - CTNNBL1 facilitates the association of CWC15 with CDC5L and is required to maintain the abundance of the Prp19 spliceosomal complex
AU - Van Maldegem, Febe
AU - Maslen, Sarah
AU - Johnson, Christopher M.
AU - Chandra, Anita
AU - Ganesh, Karuna
AU - Skehel, Mark
AU - Rada, Cristina
N1 - Funding Information: Medical Research Council [file reference number U105178806]; Leukaemia & Lymphoma Research [to F.v.M. no. 11018 and to A.C. no. 06011 & 10017]; Netherlands Organisation for Scientific Research (NWO) [825.09.003 to F.v.M.]. Funding for open access charge: Medical Research Council. Funding Information: Medical Research Council [file reference number U105178806]; Leukaemia & Lymphoma Research [to F.v.M. no. 11018 and to A.C. no. 06011 & 10017]; Netherlands Organisation for Scientific Research (NWO) [825.09.003 to F.v.M.]. Funding for open access charge: Medical Research Council. Conflict of interest statement. None declared. Publisher Copyright: © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.
PY - 2015/8/18
Y1 - 2015/8/18
N2 - In order to catalyse the splicing of messenger RNA, multiple proteins and RNA components associate and dissociate in a dynamic highly choreographed process. The Prp19 complex is a conserved essential part of the splicing machinery thought to facilitate the conformational changes the spliceosome undergoes during catalysis. Dynamic protein interactions often involve highly disordered regions that are difficult to study by structural methods. Using amine crosslinking and hydrogen-deuterium exchange coupled to mass spectrometry, we describe the architecture of the Prp19 sub-complex that contains CTNNBL1. Deficiency in CTNNBL1 leads to delayed initiation of cell division and embryonic lethality. Here we show that in vitro CTNNBL1 enhances the association of CWC15 and CDC5L, both core Prp19 complex proteins and identify an overlap in the region of CDC5L that binds either CTNNBL1 or CWC15 suggesting the two proteins might exchange places in the complex. Furthermore, in vivo, CTNNBL1 is required to maintain normal levels of the Prp19 complex and to facilitate the interaction of CWC15 with CDC5L. Our results identify a chaperone function for CTNNBL1 within the essential Prp19 complex, a function required to maintain the integrity of the complex and to support efficient splicing.
AB - In order to catalyse the splicing of messenger RNA, multiple proteins and RNA components associate and dissociate in a dynamic highly choreographed process. The Prp19 complex is a conserved essential part of the splicing machinery thought to facilitate the conformational changes the spliceosome undergoes during catalysis. Dynamic protein interactions often involve highly disordered regions that are difficult to study by structural methods. Using amine crosslinking and hydrogen-deuterium exchange coupled to mass spectrometry, we describe the architecture of the Prp19 sub-complex that contains CTNNBL1. Deficiency in CTNNBL1 leads to delayed initiation of cell division and embryonic lethality. Here we show that in vitro CTNNBL1 enhances the association of CWC15 and CDC5L, both core Prp19 complex proteins and identify an overlap in the region of CDC5L that binds either CTNNBL1 or CWC15 suggesting the two proteins might exchange places in the complex. Furthermore, in vivo, CTNNBL1 is required to maintain normal levels of the Prp19 complex and to facilitate the interaction of CWC15 with CDC5L. Our results identify a chaperone function for CTNNBL1 within the essential Prp19 complex, a function required to maintain the integrity of the complex and to support efficient splicing.
UR - http://www.scopus.com/inward/record.url?scp=84984941810&partnerID=8YFLogxK
U2 - https://doi.org/10.1093/nar/gkv643
DO - https://doi.org/10.1093/nar/gkv643
M3 - Article
C2 - 26130721
SN - 0305-1048
VL - 43
SP - 7058
EP - 7069
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 14
ER -