Antibodies specific for branched ribonucleic acids.
A chemically synthesized branched tetranucleotide, G3'p5'A [2'p5'G]3'p5'C corresponding to the consensus sequence at the branch point in introns undergoing RNA splicing, was used as a hapten to elicit antibranch antibodies. Binding assays with 32P-labeled hapten and unlabeled structurally related haptens indicated that the antibodies are highly specific for the branch structure and have some specificity for the A2'p5'G sequence at the branch point, but have essentially none for a variety of other 2'p5' or 3'p5' dinucleotides or for the linear trinucleotide G3'p5'A3'p5'C. Purification of these antibodies by binding to A2'p5'G covalently linked to Sepharose followed by covalent attachment of the purified antibodies to protein A-Sepharose has provided an adsorbent that immunospecifically retains branched oligonucleotides as well as branched introns released from RNAs during in vitro splicing.
Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260.
J D
JD
Reilly
S K
SK
Freeman
R F
RF
Melhem
R
R
Kierzek
M H
MH
Caruthers
M
M
Edmonds
T W
TW
Munns
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000915:Antibody Affinity; D000917:Antibody Formation; D000918:Antibody Specificity; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D007438:Introns; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012313:RNA / Q000276:immunology*; D012326:RNA Splicing; D011817:Rabbits; D011863:Radioimmunoassay
D000906:Antibodies; D012313:RNA