Interaction of the RNA binding fingers of Xenopus transcription factor IIIA with specific regions of 5 S ribosomal RNA.
Zinc fingers 4 to 7 of Xenopus transcription factor IIIA (TFIIIA) represent the minimal polypeptide necessary for high-affinity binding to 5 S RNA. Mutations covering the entire 5 S RNA structure have been compared for their effects on the binding affinity of full-length TFIIIA and a polypeptide consisting of fingers 4 to 7 of TFIIIA (zf4-7). In addition, ribonuclease footprinting was used to compare the binding sites of TFIIIA and zf4-7 on 5 S RNA. The consistency between the data obtained from these two approaches provided a clear indication that zinc fingers 4 to 7 of TFIIIA bind to a central core region on the 5 S RNA molecule consisting of loop B/helix II/loop A/helix V/region E. This information was used to design a truncated 75-nucleotide-long RNA molecule that retains high affinity for zf4-7. Therefore, we conclude that the specific interaction of TFIIIA with 5 S RNA can be represented by a complex formed between a four zinc finger polypeptide and a truncated 5 S RNA molecule.
McBryant
S J
SJ
Department of Molecular Biology, Scripps Research Institute, La Jolla, CA 92037, USA.
Veldhoen
N
N
Gedulin
B
B
Leresche
A
A
Foster
M P
MP
Wright
P E
PE
Romaniuk
P J
PJ
Gottesfeld
J M
JM
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D005260:Female; D018257:Helix-Loop-Helix Motifs; D007700:Kinetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009690:Nucleic Acid Conformation; D009865:Oocytes / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012341:RNA, Ribosomal, 5S / Q000737:chemistry* / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D013698:Templates, Genetic; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*; D014982:Xenopus laevis; D016335:Zinc Fingers
D017931:DNA Primers; D004268:DNA-Binding Proteins; D012341:RNA, Ribosomal, 5S; D016601:RNA-Binding Proteins; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors