Identification of the binding site on 5S rRNA for the transcription factor IIIA: proposed structure of a common binding site on 5S rRNA and on the gene.

Transcription factor IIIA (TFIIIA) employs an array of nine N-terminal zinc fingers to bind specifically to both 5S RNA and 5S DNA. The binding of TFIIIA to 5S RNA and 5S DNA was studied by using a protease footprinting technique. Brief treatment of free or bound TFIIA with trypsin or chymotrypsin generated fragments which were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Fragments retaining the N terminus of TFIIA were identified by immunoblotting with an antibody directed against the N terminus of TFIIIA. Proteolytic footprinting of TFIIIA complexed with 5S DNA derivatives reinforced other evidence that the three N-terminal zinc fingers of TFIIIA bind most tightly to 5S DNA. Proteolytic footprinting of TFIIIA in reconstituted 7S ribonucleoprotein particles revealed different patterns of trypsin sensitivity for TFIIIA bound to oocyte versus somatic 5S RNA. Trypsin cleaved TFIIIA between zinc fingers 3 and 4 more readily when the protein was bound to somatic 5S RNA than when it was bound to oocyte 5S RNA. A tryptic fragment of TFIIIA containing zinc fingers 4 through 7 remained tightly associated with somatic 5S RNA. Zinc fingers 4 through 7 may represent a tightly binding site for 5S RNA in the same sense that fingers 1 through 3 represent a tightly binding site for 5S DNA.

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Proteolytic footprinting of transcription factor TFIIIA reveals different tightly binding sites for 5S RNA and 5S DNA.

Molecular and Cellular Biology ◽

10.1128/mcb.13.9.5149 ◽

1993 ◽

Vol 13 (9) ◽

pp. 5149-5158 ◽

Cited By ~ 14

Author(s):

D F Bogenhagen

Keyword(s):

Transcription Factor ◽

Binding Site ◽

Zinc Fingers ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

5S Rna ◽

Transcription Factor Iiia ◽

Tryptic Fragment ◽

N Terminus ◽

5S Dna

Transcription factor IIIA (TFIIIA) employs an array of nine N-terminal zinc fingers to bind specifically to both 5S RNA and 5S DNA. The binding of TFIIIA to 5S RNA and 5S DNA was studied by using a protease footprinting technique. Brief treatment of free or bound TFIIA with trypsin or chymotrypsin generated fragments which were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Fragments retaining the N terminus of TFIIA were identified by immunoblotting with an antibody directed against the N terminus of TFIIIA. Proteolytic footprinting of TFIIIA complexed with 5S DNA derivatives reinforced other evidence that the three N-terminal zinc fingers of TFIIIA bind most tightly to 5S DNA. Proteolytic footprinting of TFIIIA in reconstituted 7S ribonucleoprotein particles revealed different patterns of trypsin sensitivity for TFIIIA bound to oocyte versus somatic 5S RNA. Trypsin cleaved TFIIIA between zinc fingers 3 and 4 more readily when the protein was bound to somatic 5S RNA than when it was bound to oocyte 5S RNA. A tryptic fragment of TFIIIA containing zinc fingers 4 through 7 remained tightly associated with somatic 5S RNA. Zinc fingers 4 through 7 may represent a tightly binding site for 5S RNA in the same sense that fingers 1 through 3 represent a tightly binding site for 5S DNA.

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Displacement of Xenopus transcription factor IIIA from a 5S rRNA gene by a transcribing RNA polymerase.

Molecular and Cellular Biology ◽

10.1128/mcb.11.8.3978 ◽

1991 ◽

Vol 11 (8) ◽

pp. 3978-3986 ◽

Cited By ~ 11

Author(s):

F E Campbell ◽

D R Setzer

Keyword(s):

Transcription Factor ◽

Dna Binding ◽

Rna Polymerase ◽

Internal Control ◽

Rna Polymerase Iii ◽

5S Rrna ◽

Rrna Gene ◽

Transcription Factor Iiia ◽

Polymerase Iii ◽

5S Rrna Gene

In the absence of other components of the RNA polymerase III transcription machinery, transcription factor IIIA (TFIIIA) can be displaced from both strands of its DNA-binding site (the internal control region) on the somatic-type 5S rRNA gene of Xenopus borealis during transcription elongation by bacteriophage T7 RNA polymerase, regardless of which DNA strand is transcribed. Furthermore, substantial displacement is observed after the template has been transcribed only once. Since the complete 5S rRNA transcription complex has previously been shown to remain stably bound to the gene during repeated rounds of transcription by either RNA polymerase III or bacteriophage SP6 RNA polymerase, these results indicate that a factor(s) in addition to TFIIIA is required to create a complex that will remain stably associated with the template during transcription. Thus, transcription complex stability during passage of RNA polymerase cannot be explained solely on the basis of the DNA-binding properties of TFIIIA.

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Maize TF IIIA--the first transcription factor IIIA from monocotyledons. Purification and properties.

Acta Biochimica Polonica ◽

10.18388/abp.1997_4406 ◽

1997 ◽

Vol 44 (3) ◽

pp. 579-589 ◽

Cited By ~ 3

Author(s):

E Wyszko ◽

M Radłowski ◽

S Bartkowiak ◽

M Z Barciszewska

Keyword(s):

Transcription Factor ◽

Zea Mays L ◽

Cell Extract ◽

5S Rrna ◽

Rrna Gene ◽

Maize Pollen ◽

Transcription Factor Iiia ◽

Purification And Properties ◽

5S Rrna Gene ◽

First Time

Purification and properties of transcription factor IIIA (TF IIIA) from maize pollen (Zea mays L.) are presented for the first time. The purified protein has a molecular mass of about 35 kDa and exhibits binding affinity toward both 5S rRNA and 5S rRNA gene. It also facilitates transcription of the 5S rRNA gene in a HeLa cell extract.

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The crystal structure of d(GGATGGGAG) forms an essential part of the binding site for transcription factor IIIA

Nature ◽

10.1038/322661a0 ◽

1986 ◽

Vol 322 (6080) ◽

pp. 661-664 ◽

Cited By ~ 85

Author(s):

Maxine McCall ◽

Tom Brown ◽

William N. Hunter ◽

Olga Kennard

Keyword(s):

Crystal Structure ◽

Transcription Factor ◽

Binding Site ◽

Transcription Factor Iiia

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A plant 5S rRNA mimic regulates alternative splicing of transcription factor IIIA pre‐mRNAs

The FASEB Journal ◽

10.1096/fasebj.23.1_supplement.665.4 ◽

2009 ◽

Vol 23 (S1) ◽

Author(s):

Ming Chen Hammond ◽

Andreas Wachter ◽

Ronald R. Breaker

Keyword(s):

Transcription Factor ◽

Alternative Splicing ◽

5S Rrna ◽

Transcription Factor Iiia

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Human transcription factor TFIIIC2 specifically interacts with a unique sequence in the Xenopus laevis 5S rRNA gene

Molecular and Cellular Biology ◽

10.1128/mcb.9.11.4941-4950.1989 ◽

1989 ◽

Vol 9 (11) ◽

pp. 4941-4950

Author(s):

L G Fradkin ◽

S K Yoshinaga ◽

A J Berk ◽

A Dasgupta

Keyword(s):

Transcription Factor ◽

Xenopus Laevis ◽

Binding Site ◽

Gene Transcription ◽

5S Rrna ◽

Rrna Genes ◽

Rrna Gene ◽

5S Rrna Gene ◽

5S Rrna Genes ◽

Type Gene

Transcription factor TFIIIC2 derived from human cells is required for tRNA-type gene transcription and binds with high affinity to the essential B-box promoter element of tRNA-type genes. Although 5S rRNA genes contain no homology with the tRNA-type gene B box, we show that TFIIIC2 is also required for Xenopus laevis 5S rRNA gene transcription. TFIIIC2 protected an approximately 30-base-pair (-10 to +18) region of a Xenopus 5S rRNA gene from DNase I digestion. This region, which spanned the transcription start site, included sequences that are highly conserved among eucaryotic 5S rRNA genes and have no homology with the B-box sequence of tRNA genes. Mutation of the TFIIIC2-binding site reduced transcription of the 5S rRNA gene by a factor of 10 in HeLa cell extracts. Methylation of C residues within the TFIIIC2-binding site interfered with binding of TFIIIC2. These results suggest a role of the TFIIIC2-binding sequence in 5S rRNA gene transcription. In addition, the 5S rRNA gene binding site and the tRNA-type gene B-box sequence did not compete with each other for binding to TFIIIC2 any better than did an unrelated DNA sequence, indicating that TFIIIC2 interacts with 5S rRNA genes and tRNA-type genes through separate DNA-binding domains or polypeptides.

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