Locations of contacts between individual zinc fingers of Xenopus laevis transcription factor IIIA and the internal control region of a 5S RNA gene

The differential expression of the Xenopus laevis somatic and oocyte 5S RNA genes is partially, but not solely, due to several base differences near the 5' boundary of the internal control region. A hybrid oocyte 5S gene with somatic-type base changes at +47, +53, +55, and +56 had intermediate transcriptional activity in oocyte S150 extracts. These base substitutions also resulted in increased affinity for a factor(s), other than TFIIIA, which forms a stable complex with the 5S gene.

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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.13.9.5149-5158.1993 ◽

1993 ◽

Vol 13 (9) ◽

pp. 5149-5158

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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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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Induced Fit and “Lock and Key” Recognition of 5S RNA by Zinc Fingers of Transcription Factor IIIA

Journal of Molecular Biology ◽

10.1016/j.jmb.2005.12.010 ◽

2006 ◽

Vol 357 (1) ◽

pp. 275-291 ◽

Cited By ~ 52

Author(s):

Brian M. Lee ◽

Jing Xu ◽

Bryan K. Clarkson ◽

Maria A. Martinez-Yamout ◽

H. Jane Dyson ◽

...

Keyword(s):

Transcription Factor ◽

Zinc Fingers ◽

5S Rna ◽

Induced Fit ◽

Transcription Factor Iiia

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Effect of sequence differences between somatic and oocyte 5S RNA genes on transcriptional efficiency in an oocyte S150 extract.

Molecular and Cellular Biology ◽

10.1128/mcb.8.11.5056 ◽

1988 ◽

Vol 8 (11) ◽

pp. 5056-5058 ◽

Cited By ~ 5

Author(s):

W F Reynolds

Keyword(s):

Xenopus Laevis ◽

Differential Expression ◽

Internal Control ◽

Transcriptional Activity ◽

Stable Complex ◽

5S Rna ◽

Internal Control Region ◽

5S Rna Genes ◽

Rna Genes ◽

5S Gene

The differential expression of the Xenopus laevis somatic and oocyte 5S RNA genes is partially, but not solely, due to several base differences near the 5' boundary of the internal control region. A hybrid oocyte 5S gene with somatic-type base changes at +47, +53, +55, and +56 had intermediate transcriptional activity in oocyte S150 extracts. These base substitutions also resulted in increased affinity for a factor(s), other than TFIIIA, which forms a stable complex with the 5S gene.

Download Full-text