scholarly journals The positive transcription factor of the 5S RNA gene proteolyses during direct exchange between 5S DNA sites.

1986 ◽  
Vol 103 (3) ◽  
pp. 673-681 ◽  
Author(s):  
E B Kmiec ◽  
A Worcel
Keyword(s):  
Transcription Factor ◽  
Complex Formation ◽  
5S Rna ◽  
Specific Transcription Factor ◽  
Direct Exchange ◽  
5S Dna ◽  
Dna Complex ◽  
Vector Dna

We have examined the association, dissociation, and exchange of the 5S specific transcription factor (TFIIIA) with somatic- and oocyte-type 5S DNA. The factor associates faster with somatic than with oocyte 5S DNA, and the rate of complex formation is accelerated by vector DNA. Once formed, the TFIIIA-5S DNA complex is stable for greater than 4 h in the absence of free 5S DNA, and its dissociation is identical for somatic and for oocyte 5S DNA. In the presence of free 5S DNA, the factor transfers promptly from the complex to the free 5S DNA site. Unexpectedly, the direct exchange of factor between 5S DNA sites leads to proteolysis at the C-terminal arm of TFIIIA.

The C-terminal domain of transcription factor IIIA interacts differently with different 5S RNA genes

1989 ◽  
Vol 9 (2) ◽  
pp. 499-514
Author(s):  
Y Y Xing ◽  
A Worcel
Keyword(s):  
Transcription Factor ◽  
Site Directed Mutagenesis ◽  
5S Rna ◽  
Transcription Factor Iiia ◽  
5S Dna ◽  
Type Site ◽  
Single Trace ◽  
Dna Complex ◽  
5S Rna Genes ◽  
Rna Genes

DNase I footprints and affinity measurements showed that the C-terminal arm of Xenopus transcription factor IIIA interacts differently with different Xenopus 5S DNAs, forming three distinct types of transcription factor IIIA-5S DNA complexes: a somatic type, a major-oocyte (and pseudogene) type, and a trace-oocyte type. Site-directed mutagenesis on the major-oocyte 5S gene revealed that somatic-type changes at positions 53, 55, and 56 changed the structure of the transcription factor IIIA-5S DNA complex from major-oocyte to somatic, and a single trace-oocyte change at position 56 caused the change from major-oocyte to trace-oocyte complex. We further show that the somatic-type changes are accompanied by a marked enhancement in the rate of 5S RNA transcription, and we discuss the possible biological relevance of these findings.

scholarly journals The C-terminal domain of transcription factor IIIA interacts differently with different 5S RNA genes.

1989 ◽  
Vol 9 (2) ◽  
pp. 499-514 ◽  
Author(s):  
Y Y Xing ◽  
A Worcel
Keyword(s):  
Transcription Factor ◽  
Site Directed Mutagenesis ◽  
5S Rna ◽  
Transcription Factor Iiia ◽  
5S Dna ◽  
Type Site ◽  
Single Trace ◽  
Dna Complex ◽  
5S Rna Genes ◽  
Rna Genes

DNase I footprints and affinity measurements showed that the C-terminal arm of Xenopus transcription factor IIIA interacts differently with different Xenopus 5S DNAs, forming three distinct types of transcription factor IIIA-5S DNA complexes: a somatic type, a major-oocyte (and pseudogene) type, and a trace-oocyte type. Site-directed mutagenesis on the major-oocyte 5S gene revealed that somatic-type changes at positions 53, 55, and 56 changed the structure of the transcription factor IIIA-5S DNA complex from major-oocyte to somatic, and a single trace-oocyte change at position 56 caused the change from major-oocyte to trace-oocyte complex. We further show that the somatic-type changes are accompanied by a marked enhancement in the rate of 5S RNA transcription, and we discuss the possible biological relevance of these findings.

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

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.

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

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.

Mutations in 5S DNA and 5S RNA have different effects on the binding of Xenopus transcription factor IIIA

Biochemistry ◽  
1991 ◽  
Vol 30 (9) ◽  
pp. 2495-2500 ◽  
Author(s):  
Qimin You ◽  
Nik Veldhoen ◽  
Florence Baudin ◽  
Paul J. Romaniuk
Keyword(s):  
Transcription Factor ◽  
5S Rna ◽  
Transcription Factor Iiia ◽  
5S Dna

scholarly journals Zinc Ions Inhibit the Protein–DNA Complex Formation between Cyanobacterial Transcription Factor SmtB and its Recognition DNA Sequences

2002 ◽  
Vol 43 (10) ◽  
pp. 1254-1258 ◽  
Author(s):  
Eugene Hayato Morita ◽  
Miki Wakamatsu ◽  
Koichi Uegaki ◽  
Noboru Yumoto ◽  
Yoshimasa Kyogoku ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Complex Formation ◽  
Dna Sequences ◽  
Zinc Ions ◽  
Dna Complex

scholarly journals Transcription of the Drosophila melanogaster 5S RNA gene requires an upstream promoter and four intragenic sequence elements.

1988 ◽  
Vol 8 (3) ◽  
pp. 1266-1274 ◽  
Author(s):  
S J Sharp ◽  
A D Garcia
Keyword(s):  
Transcription Factor ◽  
Drosophila Melanogaster ◽  
Transcription Factors ◽  
Transcription Initiation ◽  
Start Site ◽  
Transcription Start ◽  
5S Rna ◽  
Upstream Promoter ◽  
5S Dna ◽  
Control Regions

Linker-scanning (LS) mutations were constructed spanning the length of the Drosophila melanogaster 5S RNA gene. In vitro transcription analysis of the LS 5S DNAs revealed five transcription control regions. One control region essential for transcription initiation was identified in the 5'-flanking sequence. The major sequence determinants of this upstream promoter region were located between coordinates -39 and -26 (-30 region), but important sequences extended to the transcription start site at position 1. Since mutations in the upstream promoter did not alter the ability of 5S DNA to sequester transcription factors into a stable transcription complex, it appears that this control region involved the interaction of RNA polymerase III. Active 5S DNA transcription additionally required the four intragenic control regions (ICRs) located between coordinates 3 and 18 (ICR I), 37 and 44 (ICR II), 48 and 61 (ICR III), and 78 and 98 (ICR IV). LS mutations in each ICR decreased the ability of 5S DNA to sequester transcription factors. ICR III, ICR IV, and the spacer sequence between were similar in sequence and position to the determinant elements of the multipartite ICR of Xenopus 5S DNA. The importance of ICR III and ICR IV in transcription initiation and in sequestering transcription factors suggests the presence of an activity in D. melanogaster similar to transcription factor TFIIIA of Xenopus laevis and HeLa cells. Transcription initiation of Drosophila 5S DNA was not eliminated by LS mutations in the spacer region even though these mutations reduced the ability of the TFIIIA-like activity to bind. The previously unidentified control regions ICR I and ICR II appear to be important for the interaction of a transcription factor activity, or multiple-factor activities, distinct from the TFIIIA-like activity. The interaction of this activity with ICR I directed the selection of the transcription start site.

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