Caenorhabditis elegans ALG-1 antimorphic mutations uncover functions for Argonaute in microRNA guide strand selection and passenger strand disposal
dc.contributor.author | Zinovyeva, Anna Y. | |
dc.contributor.author | Veksler-Lublinsky, Isana | |
dc.contributor.author | Vashisht, Ajay A. | |
dc.contributor.author | Wohlschlegel, James A. | |
dc.contributor.author | Ambros, Victor R. | |
dc.date | 2022-08-11T08:10:18.000 | |
dc.date.accessioned | 2022-08-23T17:03:32Z | |
dc.date.available | 2022-08-23T17:03:32Z | |
dc.date.issued | 2015-09-22 | |
dc.date.submitted | 2015-10-01 | |
dc.identifier.citation | <p>Proc Natl Acad Sci U S A. 2015 Sep 22;112(38):E5271-80. doi: 10.1073/pnas.1506576112. Epub 2015 Sep 8. <a href="http://dx.doi.org/10.1073/pnas.1506576112" target="_blank">Link to article on publisher's site</a></p> | |
dc.identifier.issn | 0027-8424 (Linking) | |
dc.identifier.doi | 10.1073/pnas.1506576112 | |
dc.identifier.pmid | 26351692 | |
dc.identifier.uri | http://hdl.handle.net/20.500.14038/44447 | |
dc.description.abstract | MicroRNAs are regulators of gene expression whose functions are critical for normal development and physiology. We have previously characterized mutations in a Caenorhabditis elegans microRNA-specific Argonaute ALG-1 (Argonaute-like gene) that are antimorphic [alg-1(anti)]. alg-1(anti) mutants have dramatically stronger microRNA-related phenotypes than animals with a complete loss of ALG-1. ALG-1(anti) miRISC (microRNA induced silencing complex) fails to undergo a functional transition from microRNA processing to target repression. To better understand this transition, we characterized the small RNA and protein populations associated with ALG-1(anti) complexes in vivo. We extensively characterized proteins associated with wild-type and mutant ALG-1 and found that the mutant ALG-1(anti) protein fails to interact with numerous miRISC cofactors, including proteins known to be necessary for target repression. In addition, alg-1(anti) mutants dramatically overaccumulated microRNA* (passenger) strands, and immunoprecipitated ALG-1(anti) complexes contained nonstoichiometric yields of mature microRNA and microRNA* strands, with some microRNA* strands present in the ALG-1(anti) Argonaute far in excess of the corresponding mature microRNAs. We show complex and microRNA-specific defects in microRNA strand selection and microRNA* strand disposal. For certain microRNAs (for example mir-58), microRNA guide strand selection by ALG-1(anti) appeared normal, but microRNA* strand release was inefficient. For other microRNAs (such as mir-2), both the microRNA and microRNA* strands were selected as guide by ALG-1(anti), indicating a defect in normal specificity of the strand choice. Our results suggest that wild-type ALG-1 complexes recognize structural features of particular microRNAs in the context of conducting the strand selection and microRNA* ejection steps of miRISC maturation. | |
dc.language.iso | en_US | |
dc.relation | <a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=26351692&dopt=Abstract">Link to Article in PubMed</a> | |
dc.rights | <p>Publisher PDF posted as allowed by the publisher's author rights policy at http://www.pnas.org/site/aboutpnas/authorfaq.xhtml.</p> | |
dc.subject | Caenorhabditis elegans | |
dc.subject | MicroRNAs | |
dc.subject | ALG-1 | |
dc.subject | Argonaute | |
dc.subject | microRNA | |
dc.subject | microRNA* | |
dc.subject | passenger | |
dc.subject | Developmental Biology | |
dc.subject | Genetics and Genomics | |
dc.subject | Molecular Biology | |
dc.subject | Molecular Genetics | |
dc.title | Caenorhabditis elegans ALG-1 antimorphic mutations uncover functions for Argonaute in microRNA guide strand selection and passenger strand disposal | |
dc.type | Journal Article | |
dc.source.journaltitle | Proceedings of the National Academy of Sciences of the United States of America | |
dc.source.volume | 112 | |
dc.source.issue | 38 | |
dc.identifier.legacyfulltext | https://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=1038&context=pmm_pp&unstamped=1 | |
dc.identifier.legacycoverpage | https://escholarship.umassmed.edu/pmm_pp/39 | |
dc.identifier.contextkey | 7664899 | |
refterms.dateFOA | 2022-08-23T17:03:32Z | |
html.description.abstract | <p>MicroRNAs are regulators of gene expression whose functions are critical for normal development and physiology. We have previously characterized mutations in a Caenorhabditis elegans microRNA-specific Argonaute ALG-1 (Argonaute-like gene) that are antimorphic [alg-1(anti)]. alg-1(anti) mutants have dramatically stronger microRNA-related phenotypes than animals with a complete loss of ALG-1. ALG-1(anti) miRISC (microRNA induced silencing complex) fails to undergo a functional transition from microRNA processing to target repression. To better understand this transition, we characterized the small RNA and protein populations associated with ALG-1(anti) complexes in vivo. We extensively characterized proteins associated with wild-type and mutant ALG-1 and found that the mutant ALG-1(anti) protein fails to interact with numerous miRISC cofactors, including proteins known to be necessary for target repression. In addition, alg-1(anti) mutants dramatically overaccumulated microRNA* (passenger) strands, and immunoprecipitated ALG-1(anti) complexes contained nonstoichiometric yields of mature microRNA and microRNA* strands, with some microRNA* strands present in the ALG-1(anti) Argonaute far in excess of the corresponding mature microRNAs. We show complex and microRNA-specific defects in microRNA strand selection and microRNA* strand disposal. For certain microRNAs (for example mir-58), microRNA guide strand selection by ALG-1(anti) appeared normal, but microRNA* strand release was inefficient. For other microRNAs (such as mir-2), both the microRNA and microRNA* strands were selected as guide by ALG-1(anti), indicating a defect in normal specificity of the strand choice. Our results suggest that wild-type ALG-1 complexes recognize structural features of particular microRNAs in the context of conducting the strand selection and microRNA* ejection steps of miRISC maturation.</p> | |
dc.identifier.submissionpath | pmm_pp/39 | |
dc.contributor.department | RNA Therapeutics Institute | |
dc.contributor.department | Program in Molecular Medicine | |
dc.source.pages | E5271-80 |