scholarly journals CHL12, a gene essential for the fidelity of chromosome transmission in the yeast Saccharomyces cerevisiae.

Genetics ◽  
1994 ◽  
Vol 138 (4) ◽  
pp. 1067-1079 ◽  
Author(s):  
N Kouprina ◽  
E Kroll ◽  
A Kirillov ◽  
V Bannikov ◽  
V Zakharyev ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Sequence Analysis ◽  
Critical Role ◽  
Open Reading Frame ◽  
Phase Cell ◽  
Nucleotide Sequence Analysis ◽  
Protein Sequence Analysis ◽  
Dna Metabolism ◽  
Reading Frame ◽  
Factor C

Abstract We have analyzed the CHL12 gene, earlier identified in a screen for yeast mutants with increased rates of mitotic loss of chromosome III and circular centromeric plasmids. A genomic clone of CHL12 was isolated and used to map its physical position on the right arm of chromosome XIII near the ADH3 locus. Nucleotide sequence analysis of CHL12 revealed a 2.2-kb open reading frame with a 84-kD predicted protein sequence. Analysis of the sequence upstream of the CHL12 open reading frame revealed the presence of two imperfect copies of MluI motif, ACGCGT, a sequence associated with many DNA metabolism genes in yeast. Analysis of the amino acid sequence revealed that the protein contains a NTP-binding domain and shares a low degree of homology with subunits of replication factor C (RF-C). A strain containing a null allele of CHL12 was viable under standard growth conditions, and as well as original mutants exhibited an increase in the level of spontaneous mitotic recombination, slow growth and cold-sensitive phenotypes. Most of cells carrying the null chl12 mutation arrested as large budded cells with the nucleus in the neck at nonpermissive temperature that typical for cell division cycle (cdc) mutants that arrest in the cell cycle at a point either immediately preceding M phase or during S phase. Cell cycle arrest of the chl12 mutant requires the RAD9 gene. We conclude that the CHL12 gene product has critical role in DNA metabolism.

scholarly journals CTF4 (CHL15) mutants exhibit defective DNA metabolism in the yeast Saccharomyces cerevisiae.

1992 ◽  
Vol 12 (12) ◽  
pp. 5736-5747 ◽  
Author(s):  
N Kouprina ◽  
E Kroll ◽  
V Bannikov ◽  
V Bliskovsky ◽  
R Gizatullin ◽  
...  
Keyword(s):  
Cell Division ◽  
Protein Sequence ◽  
Function Analysis ◽  
Spontaneous Mutation ◽  
Terminal Region ◽  
Open Reading Frame ◽  
Nucleotide Sequence Analysis ◽  
Dna Metabolism ◽  
Reading Frame ◽  
Helix Loop Helix

We have analyzed the CTF4 (CHL15) gene, earlier identified in two screens for yeast mutants with increased rates of mitotic loss of chromosome III and artificial circular and linear chromosomes. Analysis of the segregation properties of circular minichromosomes and chromosome fragments indicated that sister chromatid loss (1:0 segregation) is the predominant mode of chromosome destabilization in ctf4 mutants, though nondisjunction events (2:0 segregation) also occur at an increased rate. Both inter- and intrachromosomal mitotic recombination levels are elevated in ctf4 mutants, whereas spontaneous mutation to canavanine resistance was not elevated. A genomic clone of CTF4 was isolated and used to map its physical and genetic positions on chromosome XVI. Nucleotide sequence analysis of CTF4 revealed a 2.8-kb open reading frame with a 105-kDa predicted protein sequence. The CTF4 DNA sequence is identical to that of POB1, characterized as a gene encoding a protein that associates in vitro with DNA polymerase alpha. At the N-terminal region of the protein sequence, zinc finger motifs which define potential DNA-binding domains were found. The C-terminal region of the predicted protein displayed similarity to sequences of regulatory proteins known as the helix-loop-helix proteins. Data on the effects of a frameshift mutation suggest that the helix-loop-helix domain is essential for CTF4 function. Analysis of sequences upstream of the CTF4 open reading frame revealed the presence of a hexamer element, ACGCGT, a sequence associated with many DNA metabolism genes in budding yeasts. Disruption of the coding sequence of CTF4 did not result in inviability, indicating that the CTF4 gene is nonessential for mitotic cell division. However, ctf4 mutants exhibit an accumulation of large budded cells with the nucleus in the neck. ctf4 rad52 double mutants grew very slowly and produced extremely high levels (50%) of inviable cell division products compared with either single mutant alone, which is consistent with a role for CTF4 in DNA metabolism.

scholarly journals The site-specific ribosomal insertion element type II of Bombyx mori (R2Bm) contains the coding sequence for a reverse transcriptase-like enzyme.

1987 ◽  
Vol 7 (6) ◽  
pp. 2221-2230 ◽  
Author(s):  
W D Burke ◽  
C C Calalang ◽  
T H Eickbush
Keyword(s):  
Amino Acid ◽  
Sequence Analysis ◽  
Transposable Elements ◽  
Reverse Transcriptase ◽  
Bombyx Mori ◽  
Open Reading Frame ◽  
Nucleotide Sequence Analysis ◽  
Type Ii ◽  
Site Specific ◽  
Reading Frame

Two classes of DNA elements interrupt a fraction of the rRNA repeats of Bombyx mori. We have analyzed by genomic blotting and sequence analysis one class of these elements which we have named R2. These elements occupy approximately 9% of the rDNA units of B. mori and appear to be homologous to the type II rDNA insertions detected in Drosophila melanogaster. Approximately 25 copies of R2 exist within the B. mori genome, of which at least 20 are located at a precise location within otherwise typical rDNA units. Nucleotide sequence analysis has revealed that the 4.2-kilobase-pair R2 element has a single large open reading frame, occupying over 82% of the total length of the element. The central region of this 1,151-amino-acid open reading frame shows homology to the reverse transcriptase enzymes found in retroviruses and certain transposable elements. Amino acid homology of this region is highest to the mobile line 1 elements of mammals, followed by the mitochondrial type II introns of fungi, and the pol gene of retroviruses. Less homology exists with transposable elements of D. melanogaster and Saccharomyces cerevisiae. Two additional regions of sequence homology between L1 and R2 elements were also found outside the reverse transcriptase region. We suggest that the R2 elements are retrotransposons that are site specific in their insertion into the genome. Such mobility would enable these elements to occupy a small fraction of the rDNA units of B. mori despite their continual elimination from the rDNA locus by sequence turnover.

The site-specific ribosomal insertion element type II of Bombyx mori (R2Bm) contains the coding sequence for a reverse transcriptase-like enzyme

1987 ◽  
Vol 7 (6) ◽  
pp. 2221-2230
Author(s):  
W D Burke ◽  
C C Calalang ◽  
T H Eickbush
Keyword(s):  
Amino Acid ◽  
Sequence Analysis ◽  
Transposable Elements ◽  
Reverse Transcriptase ◽  
Bombyx Mori ◽  
Open Reading Frame ◽  
Nucleotide Sequence Analysis ◽  
Type Ii ◽  
Site Specific ◽  
Reading Frame

Two classes of DNA elements interrupt a fraction of the rRNA repeats of Bombyx mori. We have analyzed by genomic blotting and sequence analysis one class of these elements which we have named R2. These elements occupy approximately 9% of the rDNA units of B. mori and appear to be homologous to the type II rDNA insertions detected in Drosophila melanogaster. Approximately 25 copies of R2 exist within the B. mori genome, of which at least 20 are located at a precise location within otherwise typical rDNA units. Nucleotide sequence analysis has revealed that the 4.2-kilobase-pair R2 element has a single large open reading frame, occupying over 82% of the total length of the element. The central region of this 1,151-amino-acid open reading frame shows homology to the reverse transcriptase enzymes found in retroviruses and certain transposable elements. Amino acid homology of this region is highest to the mobile line 1 elements of mammals, followed by the mitochondrial type II introns of fungi, and the pol gene of retroviruses. Less homology exists with transposable elements of D. melanogaster and Saccharomyces cerevisiae. Two additional regions of sequence homology between L1 and R2 elements were also found outside the reverse transcriptase region. We suggest that the R2 elements are retrotransposons that are site specific in their insertion into the genome. Such mobility would enable these elements to occupy a small fraction of the rDNA units of B. mori despite their continual elimination from the rDNA locus by sequence turnover.

Nucleotide sequence analysis of a zein genomic clone with a short open reading frame

Gene ◽  
1984 ◽  
Vol 28 (1) ◽  
pp. 113-118 ◽  
Author(s):  
Jean C. Kridl ◽  
Jeffrey Vieira ◽  
Irwin Rubenstein ◽  
Joachim Messing
Keyword(s):  
Sequence Analysis ◽  
Nucleotide Sequence ◽  
Genomic Clone ◽  
Open Reading Frame ◽  
Nucleotide Sequence Analysis ◽  
Reading Frame ◽  
Short Open Reading Frame

CTF4 (CHL15) mutants exhibit defective DNA metabolism in the yeast Saccharomyces cerevisiae

1992 ◽  
Vol 12 (12) ◽  
pp. 5736-5747
Author(s):  
N Kouprina ◽  
E Kroll ◽  
V Bannikov ◽  
V Bliskovsky ◽  
R Gizatullin ◽  
...  
Keyword(s):  
Cell Division ◽  
Protein Sequence ◽  
Function Analysis ◽  
Spontaneous Mutation ◽  
Terminal Region ◽  
Open Reading Frame ◽  
Nucleotide Sequence Analysis ◽  
Dna Metabolism ◽  
Reading Frame ◽  
Helix Loop Helix

We have analyzed the CTF4 (CHL15) gene, earlier identified in two screens for yeast mutants with increased rates of mitotic loss of chromosome III and artificial circular and linear chromosomes. Analysis of the segregation properties of circular minichromosomes and chromosome fragments indicated that sister chromatid loss (1:0 segregation) is the predominant mode of chromosome destabilization in ctf4 mutants, though nondisjunction events (2:0 segregation) also occur at an increased rate. Both inter- and intrachromosomal mitotic recombination levels are elevated in ctf4 mutants, whereas spontaneous mutation to canavanine resistance was not elevated. A genomic clone of CTF4 was isolated and used to map its physical and genetic positions on chromosome XVI. Nucleotide sequence analysis of CTF4 revealed a 2.8-kb open reading frame with a 105-kDa predicted protein sequence. The CTF4 DNA sequence is identical to that of POB1, characterized as a gene encoding a protein that associates in vitro with DNA polymerase alpha. At the N-terminal region of the protein sequence, zinc finger motifs which define potential DNA-binding domains were found. The C-terminal region of the predicted protein displayed similarity to sequences of regulatory proteins known as the helix-loop-helix proteins. Data on the effects of a frameshift mutation suggest that the helix-loop-helix domain is essential for CTF4 function. Analysis of sequences upstream of the CTF4 open reading frame revealed the presence of a hexamer element, ACGCGT, a sequence associated with many DNA metabolism genes in budding yeasts. Disruption of the coding sequence of CTF4 did not result in inviability, indicating that the CTF4 gene is nonessential for mitotic cell division. However, ctf4 mutants exhibit an accumulation of large budded cells with the nucleus in the neck. ctf4 rad52 double mutants grew very slowly and produced extremely high levels (50%) of inviable cell division products compared with either single mutant alone, which is consistent with a role for CTF4 in DNA metabolism.

scholarly journals Nucleotide sequence analysis and overexpression of the gene encoding a type III chloramphenicol acetyltransferase

1988 ◽  
Vol 252 (1) ◽  
pp. 173-179 ◽  
Author(s):  
I A Murray ◽  
A R Hawkins ◽  
J W Keyte ◽  
W V Shaw
Keyword(s):  
Sequence Analysis ◽  
Nucleotide Sequence ◽  
Cyclic Amp ◽  
Chloramphenicol Acetyltransferase ◽  
Open Reading Frame ◽  
Nucleotide Sequence Analysis ◽  
Calculated Molecular Mass ◽  
Reading Frame ◽  
E Coli ◽  
Type Iii

The gene catIII, encoding a type III enterobacterial chloramphenicol acetyltransferase, was cloned from the transmissible plasmid R387 into pBR322 and bacteriophage M13 mp8. Nucleotide sequence analysis of 1160 bp of DNA identified an open reading frame encoding a protein of 213 amino acid residues and a calculated molecular mass of 24965 Da. The predicted N-terminal sequence is identical with that determined by Edman degradation of chloramphenicol acetyltransferase purified from Escherichia coli harbouring R387. Sequences equivalent to the consensus motifs for initiation and rho-factor-independent termination of transcription in E. coli occur 5′ and 3′ to the catIII open reading frame. In contrast with the catI gene, present on transposon Tn9 and many enterobacterial plasmids, expression of catIII is not subject to cyclic AMP-mediated catabolite repression in vivo and there is no sequence in the 5′ non-coding DNA that resembles that deduced as the consensus for the binding of cyclic AMP receptor protein. Unique restriction-endonuclease cleavage sites were introduced adjacent to the catIII reading frame by using oligonucleotide-directed mutagenesis to facilitate insertion into E. coli expression vectors. Fully active chloramphenicol acetyltransferase represents 30-50% of the soluble protein component of cell-free extracts of E. coli containing the appropriate plasmids.

Development of a Self-Cloning System for Actinomadura verrucosospora and Identification of Polyketide Synthase Genes Essential for Production of the Angucyclic Antibiotic Pradimicin

1999 ◽  
Vol 65 (6) ◽  
pp. 2703-2709 ◽  
Author(s):  
Tohru Dairi ◽  
Yoshimitsu Hamano ◽  
Tamotsu Furumai ◽  
Toshikazu Oki
Keyword(s):  
Nucleotide Sequence ◽  
Polyketide Synthase ◽  
Selectable Marker ◽  
Plasmid Vector ◽  
Open Reading Frame ◽  
Antibiotic Biosynthesis ◽  
Nucleotide Sequence Analysis ◽  
Reading Frame ◽  
Cloning System ◽  
Open Reading Frame 1

ABSTRACT A self-cloning system for Actinomadura verrucosospora, a producer of the angucyclic antibiotic pradimicin A (PRM A), has been developed. The system is based on reproducible and reliable protoplasting and regeneration conditions for A. verrucosospora and a novel plasmid vector that consists of a replicon from a newly found Actinomadura plasmid and a selectable marker cloned from the Actinomadurastrain. The system has an efficiency of more than 105CFU/microgram of DNA. Using this system, we have cloned and identified the polyketide synthase (PKS) genes essential for PRM A biosynthesis from A. verrucosospora. Nucleotide sequence analysis of the 3.5-kb SalI-SphI fragment showed that ketosynthase subunits (open reading frame 1 [ORF1] and ORF2) of the essential PKS genes have strong similarities (59 to 89%) to those for angucyclic antibiotic biosynthesis.

scholarly journals Open reading frame 132 of Heliocoverpa armigera nucleopolyhedrovirus encodes a functional per os infectivity factor (PIF-2)

2006 ◽  
Vol 87 (9) ◽  
pp. 2563-2569 ◽  
Author(s):  
Minggang Fang ◽  
Yingchao Nie ◽  
Qian Wang ◽  
Fei Deng ◽  
Ranran Wang ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Western Blot ◽  
Blot Analysis ◽  
Post Infection ◽  
Spodoptera Exigua ◽  
Open Reading Frame ◽  
Rt Pcr ◽  
Western Blots ◽  
Reading Frame ◽  
Budded Virus

Open reading frame 132 (Ha132) of Helicoverpa armigera nucleopolyhedrovirus (HearNPV) is a homologue of per os infectivity factor 2 (pif-2) of Spodoptera exigua multiple nucleopolyhedrovirus. Sequence analysis indicated that Ha132 encoded a protein of 383 aa with a predicted molecular mass of 44.5 kDa. Alignment of HA132 and its baculovirus homologues revealed that HA132 was highly conserved among baculoviruses, with 14 absolutely conserved cysteine residues. RT-PCR indicated that Ha132 was first transcribed at 24 h post-infection. Western blot analysis showed that a 43 kDa band was detectable in HearNPV-infected HzAM1 cells from 36 h post-infection. Western blots also indicated that HA132 was a component of the occlusion-derived virus, but not of budded virus. Deletion of Ha132 from HearNPV abolished per os infectivity, but had no effect on the infectivity of the budded virus phenotype.

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