Identification of specific let-7 microRNA binding complexes in Caenorhabditis elegans.

RNA
Shih-Peng ChanFrank J Slack

Abstract

Little is known about the protein complexes required for microRNA formation and function. Here we used native gel electrophoresis to identify miRNA ribonucleoprotein complexes (miRNPs) in Caenorhabditis elegans. Our data reveal multiple distinct miRNPs that assemble on the let-7 miRNA in vitro. The formation of these complexes is affected but not abolished by alg-1 or alg-2 null mutations. The largest complex (M*) with an estimated molecular mass of >669 kDa cofractionates with the known RISC factors ALG-1, VIG-1, and TSN-1. The M* complex and two complexes, M3 and M4, with similar molecular weights of approximately 500 kDa, also assemble on all other miRNAs used in our experiments. Two smaller complexes, M1 (approximately 160 kDa) and M2 (approximately 250 kDa), assemble on the members of the let-7 miRNAs family but not lin-4 or mir-234, and their formation is highly dependent on specific sequences in the 5' seed region of let-7. Moreover, an unidentified protein, p40, which only appears in the M1 and M2 complexes, was detected by UV triggered cross-linking to let-7 but not to lin-4. The cross-linking of p40 to let-7 is also dependent on the let-7 sequence. Another unidentified protein, p13, is detected in all let-7 binding co...Continue Reading

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Citations

Apr 9, 2014·Nature Reviews. Nephrology·Cheuk-Chun Szeto, Philip K-T Li
Sep 3, 2014·International Journal of Genomics·Ayla Valinezhad OrangMina Kazemzadeh-Bavili
Jun 15, 2011·PLoS Neglected Tropical Diseases·Johnathan J DalzellAaron G Maule
Jul 6, 2014·Clinica Chimica Acta; International Journal of Clinical Chemistry·Cheuk-Chun Szeto
Mar 17, 2010·The International Journal of Biochemistry & Cell Biology·Sylvia E J Fischer
Nov 11, 2011·Wiley Interdisciplinary Reviews. RNA·Shobha Vasudevan
Jul 25, 2009·Developmental Biology·Shih-Peng Chan, Frank J Slack
Aug 18, 2009·Nature Structural & Molecular Biology·Tomoko KawamataYukihide Tomari
Nov 6, 2020·Current Opinion in Cell Biology·Himani Galagali, John K Kim

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