Publication Date:
2008-11-04
Description:
Protein-RNA interactions have critical roles in all aspects of gene expression. However, applying biochemical methods to understand such interactions in living tissues has been challenging. Here we develop a genome-wide means of mapping protein-RNA binding sites in vivo, by high-throughput sequencing of RNA isolated by crosslinking immunoprecipitation (HITS-CLIP). HITS-CLIP analysis of the neuron-specific splicing factor Nova revealed extremely reproducible RNA-binding maps in multiple mouse brains. These maps provide genome-wide in vivo biochemical footprints confirming the previous prediction that the position of Nova binding determines the outcome of alternative splicing; moreover, they are sufficiently powerful to predict Nova action de novo. HITS-CLIP revealed a large number of Nova-RNA interactions in 3' untranslated regions, leading to the discovery that Nova regulates alternative polyadenylation in the brain. HITS-CLIP, therefore, provides a robust, unbiased means to identify functional protein-RNA interactions in vivo.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2597294/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2597294/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Licatalosi, Donny D -- Mele, Aldo -- Fak, John J -- Ule, Jernej -- Kayikci, Melis -- Chi, Sung Wook -- Clark, Tyson A -- Schweitzer, Anthony C -- Blume, John E -- Wang, Xuning -- Darnell, Jennifer C -- Darnell, Robert B -- MC_U105185858/Medical Research Council/United Kingdom -- R01 NS034389/NS/NINDS NIH HHS/ -- R01 NS034389-09/NS/NINDS NIH HHS/ -- R01 NS034389-10/NS/NINDS NIH HHS/ -- R01 NS034389-11/NS/NINDS NIH HHS/ -- R01 NS034389-12/NS/NINDS NIH HHS/ -- R01 NS034389-13A1/NS/NINDS NIH HHS/ -- R01 NS040955/NS/NINDS NIH HHS/ -- R01 NS040955-05/NS/NINDS NIH HHS/ -- R01 NS34389/NS/NINDS NIH HHS/ -- Howard Hughes Medical Institute/ -- England -- Nature. 2008 Nov 27;456(7221):464-9. doi: 10.1038/nature07488. Epub 2008 Nov 2.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Laboratory of Molecular Neuro-Oncology and Howard Hughes Medical Institute, The Rockefeller University, 1230 York Avenue, New York, New York 10021, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/18978773" target="_blank"〉PubMed〈/a〉
Keywords:
Alternative Splicing/*genetics
;
Animals
;
Antigens, Neoplasm/genetics/*metabolism
;
Cell Line
;
Cross-Linking Reagents/chemistry/metabolism
;
Exons/genetics
;
Genome/*genetics
;
Genomics
;
Humans
;
Immunoprecipitation
;
Mice
;
Neocortex/*cytology
;
Neurons/*metabolism
;
Organ Specificity
;
Polyadenylation/genetics
;
RNA, Messenger/genetics/*metabolism
;
RNA-Binding Proteins/genetics/*metabolism
Print ISSN:
0028-0836
Electronic ISSN:
1476-4687
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
,
Natural Sciences in General
,
Physics
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