2016
Methyltransferase-like protein 16 binds the 3′-terminal triple helix of MALAT1 long noncoding RNA
Brown JA, Kinzig CG, DeGregorio SJ, Steitz JA. Methyltransferase-like protein 16 binds the 3′-terminal triple helix of MALAT1 long noncoding RNA. Proceedings Of The National Academy Of Sciences Of The United States Of America 2016, 113: 14013-14018. PMID: 27872311, PMCID: PMC5150381, DOI: 10.1073/pnas.1614759113.Peer-Reviewed Original ResearchConceptsGel shift assaysMetastasis-associated lung adenocarcinoma transcript 1RNA triple helicesPutative RNA methyltransferaseCompetitive gel shift assaysRNA-protein interactionsRNA stability elementAbundant nuclear proteinNative gel shift assaysRich internal loopSitu proximity ligation assayTriple helixHEK293T cell lysatesStem-loop structureProximity ligation assayT cell lysatesRNA methyltransferaseVivo UVNucleotide compositionNuclear proteinsLung adenocarcinoma transcript 1RNA immunoprecipitationStability elementMETTL16Rich tract
2015
A heterotrimer model of the complete Microprocessor complex revealed by single-molecule subunit counting
Herbert KM, Sarkar SK, Mills M, De la Herran H, Neuman KC, Steitz JA. A heterotrimer model of the complete Microprocessor complex revealed by single-molecule subunit counting. RNA 2015, 22: 175-183. PMID: 26683315, PMCID: PMC4712668, DOI: 10.1261/rna.054684.115.Peer-Reviewed Original ResearchConceptsPri-miRNA substratesMicroprocessor complexHeterotrimeric complexDeletion constructsSingle-molecule subunit countingRNA-binding proteinFull-length proteinAbsence of RNAStem-loop structureSingle-molecule photobleachingSize exclusion chromatographyPresence of RNARNaseIII enzymesPhotobleaching assaysMicroRNA biogenesisSubunit countingMammalian cellsDroshaDGCR8Fluorescent proteinHuman cellsMultiple copiesRNAProteinExact stoichiometry
2013
Phosphorylation of DGCR8 Increases Its Intracellular Stability and Induces a Progrowth miRNA Profile
Herbert KM, Pimienta G, DeGregorio SJ, Alexandrov A, Steitz JA. Phosphorylation of DGCR8 Increases Its Intracellular Stability and Induces a Progrowth miRNA Profile. Cell Reports 2013, 5: 1070-1081. PMID: 24239349, PMCID: PMC3892995, DOI: 10.1016/j.celrep.2013.10.017.Peer-Reviewed Original ResearchConceptsMicroprocessor complexRNA-binding proteinRNase III enzymeInhibition of phosphatasesStem-loop structureERK/MAPKSpecific processing activityMiRNA expression profilesExtracellular cuesMiRNA biogenesisDrosha proteinPhosphorylation sitesPrimary miRNAMammalian cellsProtein stabilityExpression profilesDGCR8Intracellular stabilityHeLa cellsCellular levelMiRNA profilesPhosphorylationMRNA levelsProteinCells
1994
The site of 3′ end formation of histone messenger RNA is a fixed distance from the downstream element recognized by the U7 snRNP.
Scharl EC, Steitz JA. The site of 3′ end formation of histone messenger RNA is a fixed distance from the downstream element recognized by the U7 snRNP. The EMBO Journal 1994, 13: 2432-2440. PMID: 8194533, PMCID: PMC395109, DOI: 10.1002/j.1460-2075.1994.tb06528.x.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBase SequenceCell NucleusCell-Free SystemCross-Linking ReagentsFurocoumarinsGuanosineHeLa CellsHistonesHumansMiceMolecular Sequence DataNucleic Acid ConformationProtein BindingRegulatory Sequences, Nucleic AcidRibonuclease HRibonucleoproteins, Small NuclearRNA Processing, Post-TranscriptionalRNA, MessengerStructure-Activity RelationshipSubstrate SpecificityConceptsHistone downstream elementU7 small nuclear ribonucleoproteinSmall nuclear ribonucleoproteinHistone messenger RNAInsertion mutantsEnd formationSite of cleavageEnd processingDownstream elementsA residuesMessenger RNAAnti-trimethylguanosine antibodyStem-loop structureWild-type substrateCross-linking studiesPremessenger RNANuclear ribonucleoproteinEnzymatic componentsNew cleavage siteNucleotides downstreamC residuesMolecular rulerCleavage siteRNAHistones
1993
A small nucleolar RNA is processed from an intron of the human gene encoding ribosomal protein S3.
Tycowski KT, Shu MD, Steitz JA. A small nucleolar RNA is processed from an intron of the human gene encoding ribosomal protein S3. Genes & Development 1993, 7: 1176-1190. PMID: 8319909, DOI: 10.1101/gad.7.7a.1176.Peer-Reviewed Original ResearchMeSH KeywordsBase SequenceCell NucleolusCell-Free SystemConserved SequenceElectrophoresis, Polyacrylamide GelHeLa CellsHumansIntronsMolecular Sequence DataNucleic Acid ConformationRestriction MappingRibosomal ProteinsRNA PrecursorsRNA Processing, Post-TranscriptionalRNA, Small NuclearSequence Analysis, RNAUracil NucleotidesConceptsSmall nucleolar RNAsNucleolar RNAsRibosomal protein S3 geneNuclear RNA polymerasesSingle-copy geneSingle primary transcriptRibosomal protein S3Secondary structure modelStem-loop structureVertebrate cellsNucleolar proteinsProtein S3Transcription signalsHuman genesRNA polymerasePrimary transcriptConserved sequencesS3 geneNucleolar snRNASame strandS3 mRNANucleotides downstreamMature endNucleolar componentsIntron 1The Cellular RNA-Binding Protein EAP Recognizes a Conserved Stem-Loop in the Epstein-Barr Virus Small RNA EBER 1
Toczyski D, Steitz J. The Cellular RNA-Binding Protein EAP Recognizes a Conserved Stem-Loop in the Epstein-Barr Virus Small RNA EBER 1. Molecular And Cellular Biology 1993, 13: 703-710. DOI: 10.1128/mcb.13.1.703-710.1993.Peer-Reviewed Original ResearchSmall RNAsCellular RNA-binding proteinsConserved stem loopRNA-binding proteinDetailed mutational analysisStem-loop structureSequence-specific mannerRNase protection experimentsDouble-stranded regionsDeletion mutantsEBER-1Mutational analysisStem loopFusion proteinProtection experimentsViral proteinsRNAProteinEpstein-Barr virus small RNAsEpstein-Barr virusBindingMutantsHerpesvirus papioNucleotidesHairpinThe cellular RNA-binding protein EAP recognizes a conserved stem-loop in the Epstein-Barr virus small RNA EBER 1.
Toczyski DP, Steitz JA. The cellular RNA-binding protein EAP recognizes a conserved stem-loop in the Epstein-Barr virus small RNA EBER 1. Molecular And Cellular Biology 1993, 13: 703-710. PMID: 8380232, PMCID: PMC358948, DOI: 10.1128/mcb.13.1.703.Peer-Reviewed Original ResearchConceptsCellular RNA-binding proteinsRNA-binding proteinDetailed mutational analysisStem-loop structureSequence-specific mannerRNase protection experimentsEpstein‐Barr virus small RNA EBER‐1Double-stranded regionsSmall RNAsDeletion mutantsEBER-1Mutational analysisFusion proteinProtection experimentsViral proteinsProteinRNAEpstein-Barr virusBindingMutantsHerpesvirus papioNucleotidesHairpinAntibodiesCells