Publication Date:
1993-08-06
Description:
Regions of Escherichia coli ribonuclease P (RNase P) RNA in proximity to a bound transfer RNA (tRNA) substrate were mapped by photoaffinity. A photoaffinity cross-linking reagent was introduced at specific sites in the interior of the native tRNA structure by modification of the 5' ends of circularly permuted tRNAs (cptRNAs). The polymerase chain reaction was used for the production of cptRNA templates. After the amplification of a segment of a tandemly duplicated tRNA gene, the cptRNA gene was transcribed in vitro to produce cptRNA. Modified cptRNAs were cross-linked to RNase P RNA, and the conjugation sites in RNase P RNA were determined by primer extension. These sites occur in phylogenetically conserved structures and sequences and identify regions of the ribozyme that form part of the tRNA binding site. The use of circularly permuted molecules to position specific modifications is applicable to the study of many inter- and intramolecular interactions.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Nolan, J M -- Burke, D H -- Pace, N R -- GM34527/GM/NIGMS NIH HHS/ -- New York, N.Y. -- Science. 1993 Aug 6;261(5122):762-5.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Biology, Indiana University, Bloomington 47405.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/7688143" target="_blank"〉PubMed〈/a〉
Keywords:
Affinity Labels
;
Base Sequence
;
Binding Sites
;
Endoribonucleases/*chemistry/metabolism
;
Escherichia coli/enzymology/genetics
;
*Escherichia coli Proteins
;
Molecular Sequence Data
;
Nucleic Acid Conformation
;
Polymerase Chain Reaction
;
RNA/chemistry/metabolism
;
RNA, Bacterial/*chemistry/genetics/metabolism
;
RNA, Catalytic/*chemistry/metabolism
;
RNA, Transfer/*chemistry/genetics/metabolism
;
Ribonuclease P
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
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