Sequence preference in RNA recognition by the nucleoporin Nup153.
The vertebrate nuclear pore protein Nup153 contains a novel RNA binding domain. This 150-amino acid region was previously found to bind preferentially to a panel of mRNAs when compared with structured RNAs, such as tRNA, U snRNA, and double-stranded RNA. The ability to broadly recognize mRNA led to the conclusion that the Nup153 RNA binding domain confers a general affinity for single-stranded RNA. Here, we have probed Nup153 RNA recognition to decipher how this unique RNA binding domain discriminates between potential targets. We first mapped the binding determinant within an RNA fragment that associates relatively robustly with the Nup153 RNA binding domain. We next designed synthetic RNA oligonucleotides to systematically delineate the features within this minimal RNA fragment that are key to Nup153 RNA-binding domain binding and demonstrated that the binding preferences of Nup153 do not reflect general preferences of an mRNA/single-stranded RNA-binding protein. We further found that the association between Nup153 and a cellular mRNA can be attributed to an interaction with specific subregions of the RNA. These results indicate that Nup153 can discriminate between mRNA and other classes of RNA transcripts due in part to direct recognition of a loose sequence motif. This information adds a new dimension to the interfaces that can contribute to recognition in mRNA export cargo selection and fate.
Ball
Jennifer R
JR
Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City, Utah 84112, USA.
Dimaano
Christian
C
Bilak
Amber
A
Kurchan
Eydiejo
E
Zundel
M Tracy
MT
Ullman
Katharine S
KS
D020816:Amino Acid Motifs; D000818:Animals; D001483:Base Sequence; D008969:Molecular Sequence Data; D028861:Nuclear Pore Complex Proteins / Q000737:chemistry* / Q000502:physiology; D009687:Nuclear Proteins / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry*; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D016914:Ribonuclease H / Q000737:chemistry; D014158:Transcription, Genetic; D029867:Xenopus Proteins / Q000737:chemistry / Q000502:physiology*; D014982:Xenopus laevis / Q000378:metabolism
D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; C519169:Nup153 protein, Xenopus; D012333:RNA, Messenger; D011994:Recombinant Proteins; D029867:Xenopus Proteins; D012313:RNA; D016914:Ribonuclease H