Nucleic Acids Research, 1988, Vol. 16, No. 22 10493-10509
© 1988
MOLECULAR BIOLOGY |
An in vitro interaction between the human U3 snRNP and 28S rRNA sequences near the 
-sarcin site
Department of Molecular Biophysics and Biochemistry, The Howard Hughes Medical Institute, Yale University Medical School 333 Cedar Street, New Haven, CT 06510, USA
*To whom correspondence should be addressed
Received August 26, 1988. Revised October 20, 1988. Accepted October 20, 1988.
Model transcripts containing mammalian pre-rRNA sequences were incubated with a HeLa cell extract, digested with T1 RNase, and immunoprecipitated with anti-(U3)RNP or control antibodies. Two overlapping fragments derived from the 3' domain of human 28S rRNA were specifically immunoprecipitated although transcripts which spanned the transcription initiation site, the ETS processing site, the 5' end of 18S, and both termini of 5.8S yielded no protected fragments. The sequence of these fragments was determined using a novel technique in which the [32P]-labeled fragment was co-fingerprinted with [3H]-labeled total transcript serving as an internal marker. The fragments immunoprecipitated derive from nucleotides 4570–4590 and 4575–4590 of human 28S and are adjacent to the 
-sarcin site. Protection most likely involves the U3 RNA since it is sensitive to pretreatment of the extract with micrococcal nuclease. Complementarity between U3 and this rRNA region is phylogenetically conserved in species ranging from human to S. cerevisiae. The possible significance of this finding is discussed.
+Present address: Department of Cell Biology, The Howard Hughes Medical Institute, One Baylor Plaza, Baylor College of Medicine, Houston, TX 77030, USA
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