Binding of human SLBP on the 3'-UTR of histone precursor H4-12 mRNA induces structural rearrangements that enable U7 snRNA anchoring.
In metazoans, cell-cycle-dependent histones are produced from poly(A)-lacking mRNAs. The 3' end of histone mRNAs is formed by an endonucleolytic cleavage of longer precursors between a conserved stem-loop structure and a purine-rich histone downstream element (HDE). The cleavage requires at least two trans-acting factors: the stem-loop binding protein (SLBP), which binds to the stem-loop and the U7 snRNP, which anchors to histone pre-mRNAs by annealing to the HDE. Using RNA structure-probing techniques, we determined the secondary structure of the 3'-untranslated region (3'-UTR) of mouse histone pre-mRNAs H4-12, H1t and H2a-614. Surprisingly, the HDE is embedded in hairpin structures and is therefore not easily accessible for U7 snRNP anchoring. Probing of the 3'-UTR in complex with SLBP revealed structural rearrangements leading to an overall opening of the structure especially at the level of the HDE. Electrophoretic mobility shift assays demonstrated that the SLBP-induced opening of HDE actually facilitates U7 snRNA anchoring on the histone H4-12 pre-mRNAs 3' end. These results suggest that initial binding of the SLBP functions in making the HDE more accessible for U7 snRNA anchoring.
Jaeger
Sophie
S
UPR 9002 Architecture et Réactivité des ARN du CNRS, Université Louis Pasteur, Institut de Biologie Moléculaire et Cellulaire, 15 rue René Descartes, F-67084 Strasbourg cedex, France.
Martin
Franck
F
Rudinger-Thirion
Joëlle
J
Giegé
Richard
R
Eriani
Gilbert
G
D020413:3' Untranslated Regions / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D006657:Histones / Q000235:genetics*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D023302:Protein Footprinting; D026721:RNA 3' End Processing; D012322:RNA Precursors / Q000737:chemistry* / Q000378:metabolism; D012342:RNA, Small Nuclear / Q000378:metabolism*; D039221:mRNA Cleavage and Polyadenylation Factors / Q000378:metabolism*
D020413:3' Untranslated Regions; D006657:Histones; D009687:Nuclear Proteins; D012322:RNA Precursors; D012342:RNA, Small Nuclear; C092663:SLBP protein, human; C405487:U7 small nuclear RNA; D039221:mRNA Cleavage and Polyadenylation Factors