Photocrosslinking of 5-iodouracil-substituted RNA and DNA to proteins.
5-Iodouracil-substituted RNA and DNA were crosslinked regiospecifically to associated proteins in yields of 70 to 94% of bound nucleic acid. Irradiation of the iodouracil chromophore with monochromatic, long-wavelength ultraviolet radiation (325 nanometers) eliminates excitation of other nucleic acid and protein chromophores. The combination of high crosslinking yields, excellent specificity, and elimination of photodamage to other chromophores represents an important advance toward the precise identification of contacts in nucleoprotein complexes.
Willis
M C
MC
Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309.
Hicke
B J
BJ
Uhlenbeck
O C
OC
Cech
T R
TR
Koch
T H
TH
D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000737:chemistry*; D036022:Capsid Proteins; D004247:DNA / Q000737:chemistry*; D016054:DNA, Protozoan / Q000737:chemistry; D004279:DNA, Viral / Q000737:chemistry; D007834:Lasers; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009698:Nucleoproteins / Q000737:chemistry*; D016813:Oxytricha; D011485:Protein Binding; D015800:Protozoan Proteins / Q000737:chemistry; D012313:RNA / Q000737:chemistry*; D012367:RNA, Viral / Q000737:chemistry; D016601:RNA-Binding Proteins; D016615:Telomere / Q000737:chemistry; D014466:Ultraviolet Rays; D014498:Uracil / Q000031:analogs & derivatives* / Q000737:chemistry
D036022:Capsid Proteins; D016054:DNA, Protozoan; D004279:DNA, Viral; D009698:Nucleoproteins; D015800:Protozoan Proteins; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D014498:Uracil; D012313:RNA; D004247:DNA; C048747:5-iodouracil