Molecular cloning and subcellular localisation of the snRNP-associated protein 69KD, a structural homologue of the proto-oncoproteins TLS and EWS with RNA and DNA-binding properties.
We recently isolated and characterised a 69 kDa protein (69KD) found associated with spliceosomal small nuclear ribonucleoproteins (snRNPs). Here, we report the molecular cloning of a cDNA encoding this protein, its nucleic acid binding properties and its subcellular localisation. Sequence analysis of the 69KD cDNA revealed: (1) that 69KD shares structural similarity with the human RNA binding proteins TLS and EWS (95% and 65% identity, respectively), the products of two genes frequently targeted by tumour-specific chromosomal translocations; (2) that 69KD contains a consensus RNA binding domain (CS-RBD) and three Arg/Gly-rich RNA binding motifs, structural features typical of many RNA binding proteins, in particular of hnRNP proteins; and (3) that 69KD contains a single putative Cys2/Cys2 zinc finger domain, a characteristic of many DNA-binding proteins. Consistent with its possession of these motifs, 69KD display a general nucleic acid binding activity, with a strong preference for guanyl and uridyl-rich RNA sequences, as well as for single-stranded and double-stranded DNA. The functional significance of this affinity for nucleic acids remains unclear. However, based on the established association of 69KD with the Sm core domain of snRNPs in vivo, these motifs might help mediate 69KD binding to snRNPs or be involved in some, as yet, unknown aspect of RNA metabolism. Consistent with both possibilities, 69KD is detected within typical snRNP containing subnuclear structures referred to as speckles, and is also more widely distributed throughout the nucleoplasm, as observed for many hnRNP proteins.
Hackl
W
W
Institut für Molekularbiologie und Tumorforschung, Philipps-Universität Marburg, Germany.
Lührmann
R
R
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D006367:HeLa Cells; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D034802:RNA-Binding Protein EWS; D034702:RNA-Binding Protein FUS; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D017381:Spliceosomes / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D034802:RNA-Binding Protein EWS; D034702:RNA-Binding Protein FUS; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear