Photocrosslinking of 5-iodouracil-substituted RNA and DNA to proteins

• Published in: Science (American Association for the Advancement of Science) Vol. 262; no. 5137; pp. 1255 - 1257
• Main Authors: WILLIS, M. C, HICKE, B. J, UHLENBECK, O. C, CECH, T. R, KOCH, T. H
• Format: Journal Article
• Language: English
• Published: Washington, DC American Association for the Advancement of Science 19.11.1993
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Base Sequence; Biological and medical sciences; Biological photosensitization; Capsid - chemistry; Capsid Proteins; Crosslinking; DNA - chemistry; Dna, deoxyribonucleoproteins; DNA, Protozoan - chemistry; DNA, Viral - chemistry; Fundamental and applied biological sciences. Psychology; Lasers; Molecular Sequence Data; Nucleic Acid Conformation; Nucleic acids; Nucleoproteins; Nucleoproteins - chemistry; Oxytricha; Photosensitization, Biological; Protein Binding; Protein crosslinking; Proteins; Protozoan Proteins - chemistry; RNA - chemistry; RNA, Viral - chemistry; RNA-Binding Proteins; Telomere - chemistry; Ultraviolet Rays; Uracil - analogs & derivatives; Uracil - chemistry; Proteins; RNA; Molecular structure; DNA; Nucleoprotein; Crosslinking; Molecular complex; Method; Photochemical reaction
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.7694369

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.