Assembly of the Yin Yang 1 transcription factor into messenger ribonucleoprotein particles requires direct RNA binding activity.
The early stages of vertebrate development depend heavily on control of maternally transcribed mRNAs that are stored for long periods in complexes termed messenger ribonucleoprotein particles (mRNPs) and utilized selectively following maturation and fertilization. The transcription factor Yin Yang 1 (YY1) is associated with cytoplasmic mRNPs in vertebrate oocytes; however, the mechanism by which any of the mRNP proteins associate with mRNA in the oocyte is unknown. Here we demonstrate the mechanism by which YY1 associates with mRNPs depends on its direct RNA binding activity. High affinity binding for U-rich single-stranded RNA and A:U RNA duplexes was observed in the nanomolar range, similar to the affinity for the cognate double-stranded DNA-binding element. Similar RNA binding affinity was observed with endogenous YY1 isolated from native mRNP complexes. In vivo expression experiments reveal epitope-tagged YY1 assembled into high molecular mass mRNPs, and assembly was blocked by microinjection of high affinity RNA substrate competitor. These findings present the first clues to how mRNPs assemble during early development.
Belak
Zachery R
ZR
Department of Anatomy and Cell Biology, College of Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
Ovsenek
Nick
N
D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D000939:Epitopes / Q000737:chemistry; D007700:Kinetics; D008954:Models, Biological; D008957:Models, Genetic; D009865:Oocytes / Q000378:metabolism; D011485:Protein Binding; D012313:RNA / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D012261:Ribonucleoproteins / Q000737:chemistry*; D013379:Substrate Specificity; D029867:Xenopus Proteins / Q000737:chemistry / Q000502:physiology*; D014982:Xenopus laevis / Q000378:metabolism; D050986:YY1 Transcription Factor / Q000737:chemistry / Q000502:physiology*
D000939:Epitopes; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; D029867:Xenopus Proteins; D050986:YY1 Transcription Factor; C494701:YY1 protein, Xenopus; C026452:messenger ribonucleoprotein; D012313:RNA