Solution structure of human U1 snRNA. Derivation of a possible three-dimensional model.
The solution structure of human U1 snRNA was investigated by using base-specific chemical probes (dimethylsulfate, carbodiimide, diethylpyrocarbonate) and RNase V1. Chemical reagents were employed under various conditions of salt and temperature and allowed information at the Watson-Crick base-pairing positions to be obtained for 66% of the U1 snRNA bases. Double-stranded or stacked regions were examined with RNase V1. The dat gained from these experiments extend and support the previous 2D model for U1snRNA. However, to elucidate some aspects of the solution data that could not be accounted for by the secondary structure model, the information gathered from structure probing was used to provide the experimental basis required to construct and to test a tertiary structure model by computer graphics modeling. As a result, U1 snRNA is shown to adopt an asymmetrical X-shape that is formed by two helical domains, each one being generated by coaxial stacking of helices at the U1 snRNA cruciform. Chemical reactivities and model building show that a few nucleotides, previously proposed to be unpaired, can form A.G and U.U non Watson-Crick base-pairs, notably in stem-loop B. The structural model we propose for regions G12 to A124 integrates stereochemical constraints and is based both on solution structure data and sequence comparisons between U1 snRNAs.
Krol
A
A
Laboratoire de Biochimie, Institut de Biologie Moléculaire et Cellulaire du CNRS, Strasbourg, France.
Westhof
E
E
Bach
M
M
Lührmann
R
R
Ebel
J P
JP
Carbon
P
P
D000477:Alkylating Agents; D001482:Base Composition; D001483:Base Sequence; D002234:Carbodiimides / Q000494:pharmacology; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D012342:RNA, Small Nuclear / Q000032:analysis / Q000235:genetics*; D012260:Ribonucleases / Q000378:metabolism; D012996:Solutions; D013463:Sulfuric Acid Esters / Q000494:pharmacology
D000477:Alkylating Agents; D002234:Carbodiimides; D012342:RNA, Small Nuclear; D012996:Solutions; D013463:Sulfuric Acid Esters; D012260:Ribonucleases; C007482:dimethyl sulfate; D004047:Diethyl Pyrocarbonate