| Content Provider |
World Health Organization (WHO)-Global Index Medicus |
| Author |
Li, Wai-ming Mackedenski, Sebastian Chan, Ching-man Barnes, Mark King, Dustin T. Van Rensburg, Gerrit Mehmood, Kashif Miller, Andrew L. Lee, Chow H. |
| Description |
Author Affiliation: Barnes M ( From the Chemistry Program, University of Northern British Columbia, Prince George, British Columbia V2N 4Z9, Canada.); van Rensburg G ( From the Chemistry Program, University of Northern British Columbia, Prince George, British Columbia V2N 4Z9, Canada.); Li WM ( From the Chemistry Program, University of Northern British Columbia, Prince George, British Columbia V2N 4Z9, Canada.); Mehmood K ( From the Chemistry Program, University of Northern British Columbia, Prince George, British Columbia V2N 4Z9, Canada.); Mackedenski S ( From the Chemistry Program, University of Northern British Columbia, Prince George, British Columbia V2N 4Z9, Canada.); Chan CM ( the Division of Life Science and The Key State Laboratory for Molecular Neuroscience, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, and.); King DT ( From the Chemistry Program, University of Northern British Columbia, Prince George, British Columbia V2N 4Z9, Canada.); Miller AL ( the Division of Life Science and The Key State Laboratory for Molecular Neuroscience, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, and the Marine Biological Laboratory, Woods Hole, Massachusetts 02543.); Lee CH ( From the Chemistry Program, University of Northern British Columbia, Prince George, British Columbia V2N 4Z9, Canada, chow.lee@unbc.ca.) |
| Abstract |
The ability of its four heterogeneous nuclear RNP-K-homology (KH) domains to physically associate with oncogenic mRNAs is a major criterion for the function of the coding region determinant-binding protein (CRD-BP). However, the particular RNA-binding role of each of the KH domains remains largely unresolved. Here, we mutated the first glycine to an aspartate in the universally conserved GXXG motif of the KH domain as an approach to investigate their role. Our results show that mutation of a single GXXG motif generally had no effect on binding, but the mutation in any two KH domains, with the exception of the combination of KH3 and KH4 domains, completely abrogated RNA binding in vitro and significantly retarded granule formation in zebrafish embryos, suggesting that any combination of at least two KH domains cooperate in tandem to bind RNA efficiently. Interestingly, we found that any single point mutation in one of the four KH domains significantly impacted CRD-BP binding to mRNAs in HeLa cells, suggesting that the dynamics of the CRD-BP-mRNA interaction vary over time in vivo. Furthermore, our results suggest that different mRNAs bind preferentially to distinct CRD-BP KH domains. The novel insights revealed in this study have important implications on the understanding of the oncogenic mechanism of CRD-BP as well as in the future design of inhibitors against CRD-BP function. |
| ISSN |
00219258 |
| e-ISSN |
1083351X |
| Journal |
Journal of Biological Chemistry |
| Issue Number |
1 |
| Volume Number |
290 |
| Language |
English |
| Publisher |
American Society for Biochemistry and Molecular Biology (United States) |
| Publisher Date |
2015-01-02 |
| Publisher Place |
United States |
| Access Restriction |
Open |
| Subject Keyword |
Open Reading Frames Proto-Oncogene Proteins C-myb Metabolism RNA, Messenger RNA, Neoplasm RNA-Binding Proteins Zebrafish Genetics Animals Antigens, CD44 Chemistry Aspartic Acid Electrophoretic Mobility Shift Assay Embryo, Nonmammalian Gene Expression Glycine HeLa Cells Mutation Protein Binding Protein Interaction Domains And Motifs Recombinant Proteins Embryology Research Support, Non-U.S. Gov't Biochemistry Molecular Biology |
| Content Type |
Text |
| Resource Type |
Article |
| Subject |
Cell Biology Biochemistry Molecular Biology |
