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Reconstitution of mammalian Cleavage Factor II involved in 3’ processing of mRNA precursors
- Publication Year :
- 2018
- Publisher :
- Cold Spring Harbor Laboratory, 2018.
-
Abstract
- Cleavage factor II (CF II) is a poorly characterized component of the multi-protein complex catalyzing 3’ cleavage and polyadenylation of mammalian mRNA precursors. We have reconstituted CF II as a heterodimer of hPcf11 and hClp1. The heterodimer is active in partially reconstituted cleavage reactions, whereas hClp1 by itself is not. Pcf11 moderately stimulates the RNA 5’ kinase activity of hClp1; the kinase activity is dispensable for RNA cleavage. CF II binds RNA with nanomolar affinity. Binding is mediated mostly by the two zinc fingers in the C-terminal region of hPcf11. RNA is bound without pronounced sequence-specificity, but extended G-rich sequences appear to be preferred. We discuss the possibility that CF II contributes to the recognition of cleavage/polyadenylation substrates through interaction with G-rich far-downstream sequence elements.
- Subjects :
- 0301 basic medicine
Multiprotein complex
Polyadenylation
Biology
Cleavage (embryo)
Article
03 medical and health sciences
RNA Precursors
Kinase activity
Molecular Biology
mRNA Cleavage and Polyadenylation Factors
Zinc finger
Messenger RNA
Binding Sites
Sequence Homology, Amino Acid
Chemistry
Phosphotransferases
Nuclear Proteins
RNA
Factor ii
030104 developmental biology
Biochemistry
Multiprotein Complexes
RNA Cleavage
Protein Multimerization
Protein Binding
Transcription Factors
Subjects
Details
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....5cfec9bd67627b07e5376570cf00dd65