The Y39A Mutation of HK022 Nun Disrupts aboxBInteraction but Preserves Termination Activity†

Biochemistry ◽  
2008 ◽  
Vol 47 (28) ◽  
pp. 7335-7341 ◽  
Author(s):  
Björn M. Burmann ◽  
Augusto Uc-Mass ◽  
Kristian Schweimer ◽  
Max E. Gottesman ◽  
Paul Rösch
Keyword(s):  
Termination Activity

scholarly journals Purification and characterization of TTFI, a factor that mediates termination of mouse ribosomal DNA transcription.

1988 ◽  
Vol 8 (9) ◽  
pp. 3891-3897 ◽  
Author(s):  
I Bartsch ◽  
C Schoneberg ◽  
I Grummt
Keyword(s):  
Sodium Dodecyl ◽  
Rrna Gene ◽  
Sequence Motif ◽  
Target Sequence ◽  
Coding Region ◽  
Transcription Termination Factor ◽  
S 100 ◽  
Pair Sequence ◽  
Termination Activity ◽  
Polymerase I

Termination of rRNA gene transcription is dependent on an 18-base-pair sequence motif, AGGTCGAC CAG AT TA NTCCG (the Sal box), which is present several times in the spacer region downstream of the 3' end of the pre-rRNA coding region. We report here the purification to molecular homogeneity of a nuclear factor which specifically interacts with the Sal box element. Addition of the isolated protein to S-100 extracts which contain low levels of the Sal box-binding protein and are therefore termination incompetent restores terminating activity, indicating that this protein is a polymerase I-specific transcription termination factor. The purified protein (termed TTFI) has a molecular weight of approximately 105,000 on sodium dodecyl sulfate-polyacrylamide gels. Mild proteolysis generates a relatively protease-resistant core which still specifically recognizes its target sequence. However, the termination activity has been lost, suggesting that the interaction with the DNA and the interaction with the transcription apparatus reside in different protein domains.

In vitro transcription termination activity of the Drosophila mitochondrial DNA-binding protein DmTTF

2005 ◽  
Vol 331 (1) ◽  
pp. 357-362 ◽  
Author(s):  
Marina Roberti ◽  
Patricio Fernandez-Silva ◽  
Paola Loguercio Polosa ◽  
Erika Fernandez-Vizarra ◽  
Francesco Bruni ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Dna Binding ◽  
Binding Protein ◽  
Transcription Termination ◽  
Dna Binding Protein ◽  
In Vitro Transcription ◽  
Termination Activity

scholarly journals Role of CIS in Replication of an IncB Plasmid

1999 ◽  
Vol 181 (9) ◽  
pp. 2765-2772 ◽  
Author(s):  
J. Praszkier ◽  
A. J. Pittard
Keyword(s):  
Partial Replication ◽  
Transcription Terminator ◽  
Repa Protein ◽  
Initiation Of Replication ◽  
Position Sequence ◽  
Dna Elements ◽  
Termination Activity ◽  
Replication Activity

ABSTRACT Replication of the IncB plasmid pMU720 requires the synthesis of the cis-acting RepA protein and the presence of two DNA elements, ori and CIS. CIS is the 166-bp sequence separating the RepA coding sequence from ori. To investigate how this organization of the pMU720 replicon contributes to the mechanism of initiation of replication, mutations in the sequence and/or the length of CIS were introduced into theCIS region and their effects on the efficiency of replication of the pMU720 replicon in vivo was determined. TheCIS region was found to be composed of two domains. TherepA-proximal domain, which showed strong transcription termination activity, could be replaced by equivalent sequences from I-complex and IncL/M plasmids, whose replicons are organized in the same fashion as pMU720. Replacement by a trpA transcription terminator afforded only partial replication activity. TherepA-distal domain was shown to be a spacer whose role was to position sequence(s) within ori on the correct face of the DNA helix vis-à-vis the repA-proximal portion ofCIS. A model for the loading of RepA protein ontoori is discussed.

scholarly journals Two classes of EF1-family translational GTPases encoded by giant viruses

2019 ◽  
Vol 47 (11) ◽  
pp. 5761-5776
Author(s):  
Alexandra Zinoviev ◽  
Kazushige Kuroha ◽  
Tatyana V Pestova ◽  
Christopher U T Hellen
Keyword(s):  
Functional Characterization ◽  
Elongation Factor ◽  
Multiple Sequence ◽  
Translation Elongation ◽  
Trna Synthetases ◽  
Translation Apparatus ◽  
Giant Viruses ◽  
Termination Activity ◽  
A Site

Abstract Giant viruses have extraordinarily large dsDNA genomes, and exceptionally, they encode various components of the translation apparatus, including tRNAs, aminoacyl-tRNA synthetases and translation factors. Here, we focused on the elongation factor 1 (EF1) family of viral translational GTPases (trGTPases), using computational and functional approaches to shed light on their functions. Multiple sequence alignment indicated that these trGTPases clustered into two groups epitomized by members of Mimiviridae and Marseilleviridae, respectively. trGTPases in the first group were more closely related to GTP-binding protein 1 (GTPBP1), whereas trGTPases in the second group were closer to eEF1A, eRF3 and Hbs1. Functional characterization of representative GTPBP1-like trGTPases (encoded by Hirudovirus, Catovirus and Moumouvirus) using in vitro reconstitution revealed that they possess eEF1A-like activity and can deliver cognate aa-tRNAs to the ribosomal A site during translation elongation. By contrast, representative eEF1A/eRF3/Hbs1-like viral trGTPases, encoded by Marseillevirus and Lausannevirus, have eRF3-like termination activity and stimulate peptide release by eRF1. Our analysis identified specific aspects of the functioning of these viral trGTPases with eRF1 of human, amoebal and Marseillevirus origin.

Transcription terminator-like element within a Saccharomyces cerevisiae promoter region

1986 ◽  
Vol 6 (4) ◽  
pp. 1095-1101
Author(s):  
J G Yarger ◽  
G Armilei ◽  
M C Gorman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Promoter Region ◽  
Base Pairs ◽  
Transcription Terminator ◽  
Consensus Sequences ◽  
Physiological Relevance ◽  
Bona Fide ◽  
Dna Fragment ◽  
Termination Activity

We analyzed a cloned fragment of the yeast URA3 promoter region that contains a sequence of DNA capable of functioning as a highly efficient transcription terminator. BAL 31 deletions have shown the signal for the transcription termination activity is less than or equal to 110 base pairs and resides between bases 45 and 155 upstream of the URA3 primary ATG codon at base 227. In our in vivo assay system, the DNA fragment is able to terminate transcripts very efficiently in either orientation. The terminated transcripts bind to oligodeoxythymidylate cellulose columns and promote the synthesis of full-length cDNAs, suggesting that the transcripts are polyadenylated. The 110-base-pair region contains no sequence resembling terminator consensus sequences described by Zaret and Sherman (K.S. Zaret and F. Sherman, Cell, 28:563-573, 1982) or Henikoff and Cohen (S. Henikoff and E.H. Cohen, Mol. Cell. Biol., 4:1515-1520, 1984). We discuss the possible physiological relevance of this sequence to bona fide termination of transcription and to URA3 regulation in Saccharomyces cerevisiae.

scholarly journals Factor-dependent archaeal transcription termination

2017 ◽  
Vol 114 (33) ◽  
pp. E6767-E6773 ◽  
Author(s):  
Julie E. Walker ◽  
Olivia Luyties ◽  
Thomas J. Santangelo
Keyword(s):  
Transcription Factors ◽  
Dna Sequences ◽  
Transcription Termination ◽  
Polymerase Activity ◽  
Elongation Complex ◽  
Rna Sequences ◽  
Nascent Rna ◽  
Termination Activity ◽  
Archaeal Transcription

RNA polymerase activity is regulated by nascent RNA sequences, DNA template sequences, and conserved transcription factors. Transcription factors promoting initiation and elongation have been characterized in each domain, but transcription termination factors have been identified only in bacteria and eukarya. Here we describe euryarchaeal termination activity (Eta), the first archaeal termination factor capable of disrupting the transcription elongation complex (TEC), detail the rate of and requirements for Eta-mediated transcription termination, and describe a role for Eta in transcription termination in vivo. Eta-mediated transcription termination is energy-dependent, requires upstream DNA sequences, and disrupts TECs to release the nascent RNA to solution. Deletion of TK0566 (encoding Eta) is possible, but results in slow growth and renders cells sensitive to DNA damaging agents. Our results suggest that the mechanisms used by termination factors in archaea, eukarya, and bacteria to disrupt the TEC may be conserved, and that Eta stimulates release of stalled or arrested TECs.

scholarly journals Transcription termination signals in the nin region of bacteriophage lambda: identification of Rho-dependent termination regions.

Genetics ◽  
1995 ◽  
Vol 140 (3) ◽  
pp. 875-887 ◽  
Author(s):  
S W Cheng ◽  
D L Court ◽  
D I Friedman
Keyword(s):  
Transcription Termination ◽  
Bacteriophage Lambda ◽  
Point Mutations ◽  
Open Reading Frames ◽  
Deletion Analysis ◽  
Testing System ◽  
Transcription Antitermination ◽  
Termination Activity ◽  
Phage Growth ◽  
Reading Frames

Abstract The approximately 3-kb nin region of bacteriophage lambda, located between genes P and Q contains transcription termination signals as well as 10 open reading frames. Deletions in the nin region frees phage growth from dependence on the lambda-encoded N-transcription antitermination system, conferring a Nin phenotype (N-independence). A subregion of nin, roc, is defined by a 1.9-kb deletion (delta roc) which partially frees lambda growth from the requirement for N antitermination. The roc region has strong transcription termination activity as assayed by a plasmid-based terminator testing system. We report the following features of the roc region: the biologically significant terminators in the roc region are Rho dependent, deletion analysis located the biologically significant termination signals to a 1.2 kb-segment of roc, and analysis of other deletions and point mutations in the roc region suggested at least two biologically significant regions of termination, tR3 (extending from bp 42020 to 42231) and tR4 (extending from bp 42630 to 42825).

scholarly journals Methylation of Bacterial Release Factors RF1 and RF2 Is Required for Normal Translation Termination in Vivo

2007 ◽  
Vol 282 (49) ◽  
pp. 35638-35645 ◽  
Author(s):  
Liliana Mora ◽  
Valérie Heurgué-Hamard ◽  
Miklos de Zamaroczy ◽  
Stephanie Kervestin ◽  
Richard H. Buckingham
Keyword(s):  
Translation Termination ◽  
Carbon Sources ◽  
Ribosomal Subunit ◽  
Optimal Growth ◽  
Release Factors ◽  
Rich Media ◽  
Termination Activity ◽  
Hydrolysis Of

Bacterial release factors RF1 and RF2 are methylated on the Gln residue of a universally conserved tripeptide motif GGQ, which interacts with the peptidyl transferase center of the large ribosomal subunit, triggering hydrolysis of the ester bond in peptidyl-tRNA and releasing the newly synthesized polypeptide from the ribosome. In vitro experiments have shown that the activity of RF2 is stimulated by Gln methylation. The viability of Escherichia coli K12 strains depends on the integrity of the release factor methyltransferase PrmC, because K12 strains are partially deficient in RF2 activity due to the presence of a Thr residue at position 246 instead of Ala. Here, we study in vivo RF1 and RF2 activity at termination codons in competition with programmed frameshifting and the effect of the Ala-246 → Thr mutation. PrmC inactivation reduces the specific termination activity of RF1 and RF2(Ala-246) by ∼3- to 4-fold. The mutation Ala-246 → Thr in RF2 reduces the termination activity in cells ∼5-fold. After correction for the decrease in level of RF2 due to the autocontrol of RF2 synthesis, the mutation Ala-246 → Thr reduced RF2 termination activity by ∼10-fold at UGA codons and UAA codons. PrmC inactivation had no effect on cell growth in rich media but reduced growth considerably on poor carbon sources. This suggests that the expression of some genes needed for optimal growth under such conditions can become growth limiting as a result of inefficient translation termination.

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