title: RRM proteins interacting with the cap region of topoisomerase I.
abstract: RNA recognition motif (RRM) domains bind both nucleic acids and proteins. Several proteins that contain two closely spaced RRM domains were previously found in protein complexes formed by the cap region of human topoisomerase I, a nuclear enzyme responsible for DNA relaxation or phosphorylation of SR splicing proteins. To obtain molecular insight into specific interactions between the RRM proteins and the cap region of topo I we examined their binary interactions using the yeast two-hybrid system. The interactions were established for hnRNP A1, p54(nrb) and SF2/ASF, but not for hnRNP L or HuR. To identify the amino acid pattern responsible for binding, experimental mutagenesis was employed and computational modelling of these processes was carried out. These studies revealed that two RRM domains and six residues of the consensus sequence are required for the binding to the cap region. On the basis of the above data, a structural model for the hnRNP A1-topoisomerase I complex was proposed. The main component of the hnRNP A1 binding site is a hydrophobic pocket on the beta-surface of the first RRM domain, similar to that described for Y14 protein interacting with Mago. We demonstrated that the interaction between RRM domains and the cap region was important for the kinase reaction catalyzed by topoisomerase I. Together with the previously described inhibitory effect of RRM domains of SF2/ASF on DNA cleavage, the above suggests that the binding of RRM proteins could regulate the activity of topoisomerase I.
authors.affiliation: Institute of Biochemistry, Faculty of Biology, Warsaw University, Miecznikowa 1, 02-096 Warsaw, Poland.
authors.forename: Agata M
authors.initials: AM
authors.lastname: Trzcińska-Daneluti
authors.forename: Adam
authors.initials: A
authors.lastname: Górecki
authors.forename: Alicja
authors.initials: A
authors.lastname: Czubaty
authors.forename: Barbara
authors.initials: B
authors.lastname: Kowalska-Loth
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Girstun
authors.forename: Magdalena
authors.initials: M
authors.lastname: Murawska
authors.forename: Bogdan
authors.initials: B
authors.lastname: Lesyng
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Staroń
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004264:DNA Topoisomerases, Type I / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D034441:Heterogeneous-Nuclear Ribonucleoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D020798:Two-Hybrid System Techniques
chemical_list: D004268:DNA-Binding Proteins; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D004264:DNA Topoisomerases, Type I