Characterization of the RNA binding properties of transcription factor IIIA ofXenopus laevisoocytes
Open Access
- 1 January 1985
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 13 (14) , 5369-5387
- https://doi.org/10.1093/nar/13.14.5369
Abstract
A nitrocellulose filter binding assay has been developed to study the interaction of Xenopus transcription factor IIIA with 5S RNA. The protein binds Xenopus oocyte 55 RNA with an association constant of 1.4 × 109 M−1 at 0.1 M salt, pH 7.5 at 20°C. TF IIIA binds wheat germ 5S RNA with a two-fold higher affinity, E. coli 5S RNA with a four-fold weaker affinity, and has a barely detectable interaction with yeast tRNAphe. The preference for binding eukaryotic 5S RNA is enhanced in competition assays. The homologous reconstituted complex contains one molecule each of protein and 5S RNA and is indistinguishable from native 7S RNP in mobility on non-denaturing polyacrylamide gels. The conformation of the RNA in reconstituted particles is identical to the conformation of RNA in native 7S RNP. Further analysis of the homologous interaction reveals that complex formation is favoured both by enthalpy and entropy. The 5S RNA binding activity has a broad pH optimum spanning pH 6.0 to pH 8.0. Determination of the salt dependence of Ka reveals that as many as 5 lysine-phosphate type ionic bonds may be formed in the homologous complex. Approximately 68% of the free energy of complex formation is contributed by non-electrostatic interactions between TF IIIA and Xenopus 5S RNA.Keywords
This publication has 32 references indexed in Scilit:
- Xenopus 5S gene transcription factor, TFIIIA: Characterization of a cDNA clone and measurement of RNA levels throughout developmentCell, 1984
- 5S RNA structure and interaction with transcription factor A. 1. Ribonuclease probe of the structure of 5S RNA from Xenopus laevis oocytesBiochemistry, 1984
- 5S RNA structure and interaction with transcription factor A. 2. Ribonuclease probe of the 7S particle from Xenopus laevis immature oocytes and RNA exchange properties of the 7S particleBiochemistry, 1984
- Isolation and characterization of a 7 S RNP particle from mature Xenopus laevis oocytesFEBS Letters, 1983
- Sequence-specific interaction of R17 coat protein with its ribonucleic acid binding siteBiochemistry, 1983
- Specific interaction of a purified transcription factor with an internal control region of 5S RNA genesCell, 1980
- A control region in the center of the 5S RNA gene directs specific initiation of transcription: II. The 3′ border of the regionCell, 1980
- A control region in the center of the 5S RNA gene directs specific initiation of transcription: I. The 5′ border of the regionCell, 1980
- Structure and macromolecular interactions of 5S RNA. 2. Parameters for the interaction of ribosomal proteins L5, L18, and L25 with 5S RNA from Escherichia coliBiochemistry, 1978
- Ion effects on ligand-nucleic acid interactionsJournal of Molecular Biology, 1976