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A model for DHX15 mediated disassembly of A-complex spliceosomes

RNA. 2022-01; 
Hannah M Maul-Newby, Angela N Amorello, Turvi Sharma, John H Kim, Matthew S Modena, Beth E Prichard, Melissa S Jurica
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Catalog Antibody … These results lead us to hypothesize that DHX15 plays a quality control function in U2 … For other co-IP's, 3 µg of anti-V5 antibody (GenScript, A01724) was added to 2 mM magnesium … Get A Quote

摘要

A critical step of pre-mRNA splicing is the recruitment of U2 snRNP to the branch point sequence of an intron. U2 snRNP conformation changes extensively during branch helix formation, and several RNA-dependent ATPases are implicated in the process. However, the molecular mechanisms involved remain to be fully dissected. We took advantage of the differential nucleotide triphosphates requirements for DExD/H-box enzymes to probe their contributions to in vitro spliceosome assembly. Both ATP and GTP hydrolysis support the formation of A-complex, indicating the activity of a DEAH-enzyme because DEAD-enzymes are selective for ATP. We immunodepleted DHX15 to assess its involvement, and although splicing efficiency dec... More

关键词

A-complex, DExH NTPase, DHX15, PRP43, U2 snRNP, spliceosome, splicing