The role of zinc finger linkers in p43 and TFIIIA binding to 5S rRNA and DNA.
Transcription factor IIIA (TFIIIA) and p43 zinc finger protein form distinct complexes with 5S ribosomal RNA in Xenopus oocytes. Additionally, TFIIIA binds the internal promoter of the 5S RNA gene and supports assembly of a transcription initiation complex. Both proteins have nine tandemly repeated zinc fingers with almost identical linker lengths between corresponding fingers, yet p43 has no detectable affinity for the 5S RNA gene. TFIIIA zinc fingers 1-3 are connected by highly conserved linkers, first identified in the Drosophila protein Krüppel, that are found in many DNA binding zinc finger proteins. To understand the role of these linkers in RNA and DNA binding we exchanged three TFIIIA linker amino acids with the equivalent amino acids from p43. The major effect of linker substitution is a 50-fold reduction in DNA specificity, concomitant with an 8-fold reduction in affinity. N-Terminal zinc fingers from either TFIIIA or p43 bind to multiple specific sites on 5S RNA that are resistant to competition by tRNA or poly(rA). This mode of RNA binding is unaffected by linker substitution. These data suggest that zinc finger linkers significantly facilitate the specificity of DNA binding.
Ryan
R F
RF
Department of Microbiology and Immunology, Kimmel Cancer Institute, Thomas Jefferson University, 233 South 10th Street, Philadelphia, PA 19107, USA.
Darby
M K
MK
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism*; D017124:Conserved Sequence; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D024101:Mitochondrial Proteins; D009865:Oocytes; D010444:Peptide Elongation Factor Tu / Q000235:genetics / Q000378:metabolism*; D012341:RNA, Ribosomal, 5S / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis; D016335:Zinc Fingers
D000596:Amino Acids; D000951:Antigens, Neoplasm; D004268:DNA-Binding Proteins; D024101:Mitochondrial Proteins; D012341:RNA, Ribosomal, 5S; D011993:Recombinant Fusion Proteins; C091648:TUFM protein, human; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors; D004247:DNA; D010444:Peptide Elongation Factor Tu