[General method of isolation and analysis of polynucleotide fragments cross-linked with proteins].
A fragment of 16S RNA, cross-linked to S7 protein by UV irradiation of the 30S subunit of E. coli ribosome, was obtained by the action of T1 ribonuclease on the irradiated nucleoprotein. The digest was treated with polynucleotide kinase in the presence of [gamma-32P]ATP and the S7-cross-linked oligonucleotides were isolated. An individual oligonucleotide attached to S7 protein was obtained after proteinase treatment of the respective spot followed by electrophoresis. Sequencing of this oligonucleotide established its structure as 1233-1240 fragment of 16S RNA, the U1239 residue being the site of the S7 cross-linking. The developed general approach can be used for localizing protein - cross-linked residues in polynucleotides, whatever is the procedure employed for cross-linking.
Abdurashidova
G G
GG
Tsvetkova
E A
EA
Budovskiĭ
E I
EI
D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000032:analysis; D009841:Oligonucleotides / Q000032:analysis / Q000302:isolation & purification*; D012329:RNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D012335:RNA, Ribosomal / Q000032:analysis / Q000302:isolation & purification*; D012261:Ribonucleoproteins / Q000032:analysis / Q000302:isolation & purification*; D012269:Ribosomal Proteins / Q000032:analysis / Q000302:isolation & purification*
D009841:Oligonucleotides; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D012261:Ribonucleoproteins; D012269:Ribosomal Proteins; C037442:ribosomal protein S7
Obshchiĭ metod vydeleniia i analiza sshitykh s belkom fragmentov polinukleotidov.