Sequence-specific binding of the N-terminal three-finger fragment of Xenopus transcription factor IIIA to the internal control region of a 5S RNA gene

An N-terminal fragment of Xenopus TFIIIA, containing domains 1–3, (TF 3), was expressed in E. coli. High yields of recombinant zinc finger protein was isolated, and its DNA binding activity for the internal control region (ICR) of the Xenopus 5S RNA gene, was demonstrated by bandshift experiments an...

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Bibliographic Details
Published in:FEBS letters 1991-04, Vol.281 (1), p.181-184
Main Authors: Christensen, Jorgen Holst, Hansen, Peter Kamp, Lillelund, Ove, Thøgersen, Hans Christian
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Biological and medical sciences
Cloning, Molecular
Deoxyribonuclease I
DNA, Ribosomal - genetics
DNA, Ribosomal - isolation & purification
DNA, Ribosomal - metabolism
Domain cooperativity
Domain structure
Escherichia coli - genetics
Footprinting
Fundamental and applied biological sciences. Psychology
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Plasmids
Protein Binding
Restriction Mapping
RNA, Ribosomal, 5S - genetics
Transcription factor IIIA
Transcription Factor TFIIIA
Transcription Factors - metabolism
Transcription. Transcription factor. Splicing. Rna processing
Xenopus
Zinc finger
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Summary:An N-terminal fragment of Xenopus TFIIIA, containing domains 1–3, (TF 3), was expressed in E. coli. High yields of recombinant zinc finger protein was isolated, and its DNA binding activity for the internal control region (ICR) of the Xenopus 5S RNA gene, was demonstrated by bandshift experiments and DNase I footprinting analysis. TF 3 protects 20 bp of ICR against DNase I digestion. The limits of protection are from +77 to +96 on both coding and noncoding strand. This protection pattern is identical to the protection pattern obtained with TFIIIA in the overlapping region, showing that the 3-finger fragment accounts fully for the protein-DNA interactions in TFIIIA-5S RNA gene over this region.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(91)80388-J