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PMID: 1341195 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

Molecular and cellular biology ·Vol. 12 ·No. 12 ·1992-12-00 ·Pages 5736-47

Kouprina N, Kroll E, Bannikov V, Bliskovsky V, Gizatullin R, Kirillov A, Shestopalov B, Zakharyev V, Hieter P, Spencer F

Abstract

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.

MeSH Terms
Amino Acid Sequence Base Sequence Chromosome Mapping Cloning, Molecular DNA, Fungal/metabolism DNA-Binding Proteins/chemistry,genetics,metabolism Fungal Proteins/chemistry,genetics,metabolism Genes, Fungal Genetic Complementation Test Kinetics Molecular Sequence Data Mutation Proteins Restriction Mapping Saccharomyces cerevisiae/genetics,growth & development,metabolism Saccharomyces cerevisiae Proteins
Chemicals
CTF4 protein, S cerevisiae DNA, Fungal DNA-Binding Proteins Fungal Proteins Proteins Saccharomyces cerevisiae Proteins
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Kouprina N
Institute of Cytology, Academy of Sciences of Russia, St. Petersburg.
Kroll E
Bannikov V
Bliskovsky V
Gizatullin R
Kirillov A
Shestopalov B
Zakharyev V
Hieter P
Spencer F
References (52)
52 references, click to expand
  1. The CHL 1 (CTF 1) gene product of Saccharomyces cerevisiae is important for chromosome transmission and normal cell cycle progression in G2/M.
    EMBO J. 1990 Dec;9(13):4347-58 PMID: 2265610
  2. Promoters, activator proteins, and the mechanism of transcriptional initiation in yeast.
    Cell. 1987 May 8;49(3):295-7 PMID: 2882858
  3. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  4. Flow cytometric determinations of cellular substances in algae, bacteria, moulds and yeasts.
    Antonie Van Leeuwenhoek. 1978;44(3-4):269-82 PMID: 378116
  5. The repair of double-strand breaks in the nuclear DNA of Saccharomyces cerevisiae and its genetic control.
    Mol Gen Genet. 1976 Jan 16;143(2):119-29 PMID: 765749
  6. Evidence that POB1, a Saccharomyces cerevisiae protein that binds to DNA polymerase alpha, acts in DNA metabolism in vivo.
    Mol Cell Biol. 1992 Dec;12(12):5724-35 PMID: 1448101
  7. Activation of replication origins within yeast chromosomes.
    Annu Rev Cell Biol. 1991;7:375-402 PMID: 1809350
  8. Zinc fingers, zinc clusters, and zinc twists in DNA-binding protein domains.
    Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):999-1003 PMID: 1846973
  9. Zinc finger motif for single-stranded nucleic acids? Investigations by nuclear magnetic resonance.
    J Cell Biochem. 1991 Jan;45(1):41-8 PMID: 2005183
  10. Positional mapping of genes by chromosome blotting and chromosome fragmentation.
    Methods Enzymol. 1991;194:57-77 PMID: 2005810
  11. Coordination of expression of DNA synthesis genes in budding yeast by a cell-cycle regulated trans factor.
    Nature. 1991 Mar 21;350(6315):247-50 PMID: 2005980
  12. Centromere structure and function in budding and fission yeasts.
    New Biol. 1990 Jan;2(1):10-9 PMID: 2078550
  13. Mutations that disrupt DNA binding and dimer formation in the E47 helix-loop-helix protein map to distinct domains.
    Proc Natl Acad Sci U S A. 1990 Jun;87(12):4722-6 PMID: 2112746
  14. KAR3, a kinesin-related gene required for yeast nuclear fusion.
    Cell. 1990 Mar 23;60(6):1029-41 PMID: 2138512
  15. The protein Id: a negative regulator of helix-loop-helix DNA binding proteins.
    Cell. 1990 Apr 6;61(1):49-59 PMID: 2156629
  16. A third essential DNA polymerase in S. cerevisiae.
    Cell. 1990 Sep 21;62(6):1143-51 PMID: 2169349
  17. Yeast centromere binding protein CBF1, of the helix-loop-helix protein family, is required for chromosome stability and methionine prototrophy.
    Cell. 1990 May 4;61(3):437-46 PMID: 2185892
  18. Mitotic transmission of artificial chromosomes in cdc mutants of the yeast, Saccharomyces cerevisiae.
    Genetics. 1990 Aug;125(4):763-74 PMID: 2204582
  19. Chromosome instability mutants of Saccharomyces cerevisiae that are defective in microtubule-mediated processes.
    Mol Cell Biol. 1990 Jan;10(1):223-34 PMID: 2403635
  20. Mitotic chromosome transmission fidelity mutants in Saccharomyces cerevisiae.
    Genetics. 1990 Feb;124(2):237-49 PMID: 2407610
  21. A new DNA binding and dimerization motif in immunoglobulin enhancer binding, daughterless, MyoD, and myc proteins.
    Cell. 1989 Mar 10;56(5):777-83 PMID: 2493990
  22. Interactions between heterologous helix-loop-helix proteins generate complexes that bind specifically to a common DNA sequence.
    Cell. 1989 Aug 11;58(3):537-44 PMID: 2503252
  23. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
    Genetics. 1989 May;122(1):19-27 PMID: 2659436
  24. Structure and function of telomeres.
    Annu Rev Genet. 1989;23:579-604 PMID: 2694944
  25. RAD3 protein of Saccharomyces cerevisiae is a DNA helicase.
    Proc Natl Acad Sci U S A. 1987 Dec;84(24):8951-5 PMID: 2827162
  26. The stability of chromosomes in yeast.
    Curr Genet. 1987;11(6-7):435-43 PMID: 2836079
  27. Characterization of a mutation in yeast causing nonrandom chromosome loss during mitosis.
    Genetics. 1978 Apr;88(4 Pt 1):651-71 PMID: 17176533
  28. Deletion analysis of GAL4 defines two transcriptional activating segments.
    Cell. 1987 Mar 13;48(5):847-53 PMID: 3028647
  29. Genetic control of chromosome stability in the yeast Saccharomyces cerevisiae.
    Yeast. 1988 Dec;4(4):257-69 PMID: 3064490
  30. Amino acid sequence template useful for alpha-helix-turn-alpha-helix prediction.
    FEBS Lett. 1988 Jun 6;233(1):105-8 PMID: 3289965
  31. The CDC8 transcript is cell cycle regulated in yeast and is expressed coordinately with CDC9 and CDC21 at a point preceding histone transcription.
    Exp Cell Res. 1987 Jul;171(1):223-31 PMID: 3305044
  32. Normal stoichiometry of histone dimer sets is necessary for high fidelity of mitotic chromosome transmission.
    Cell. 1986 Jan 17;44(1):43-52 PMID: 3510079
  33. Altered fidelity of mitotic chromosome transmission in cell cycle mutants of S. cerevisiae.
    Genetics. 1985 Jul;110(3):381-95 PMID: 3894160
  34. Mitotic stability of yeast chromosomes: a colony color assay that measures nondisjunction and chromosome loss.
    Cell. 1985 Feb;40(2):381-92 PMID: 3967296
  35. Sequence of a yeast DNA fragment containing a chromosomal replicator and the TRP1 gene.
    Gene. 1980 Jul;10(2):157-66 PMID: 6248420
  36. One-step gene disruption in yeast.
    Methods Enzymol. 1983;101:202-11 PMID: 6310324
  37. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  38. Isolation of the beta-tubulin gene from yeast and demonstration of its essential function in vivo.
    Cell. 1983 May;33(1):211-9 PMID: 6380751
  39. Isolation of a yeast centromere and construction of functional small circular chromosomes.
    Nature. 1980 Oct 9;287(5782):504-9 PMID: 6999364
  40. Mitotic chromosome loss in a radiation-sensitive strain of the yeast Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1981 Sep;78(9):5778-82 PMID: 7029545
  41. Structure and function of the Saccharomyces cerevisiae CDC2 gene encoding the large subunit of DNA polymerase III.
    EMBO J. 1989 Jun;8(6):1849-54 PMID: 2670563
  42. Structure and expression of the murine L-myc gene.
    EMBO J. 1987 Nov;6(11):3359-66 PMID: 2828024
  43. MSS18, a yeast nuclear gene involved in the splicing of intron aI5 beta of the mitochondrial cox1 transcript.
    EMBO J. 1988 May;7(5):1455-64 PMID: 2842150
  44. Rapid and sensitive protein similarity searches.
    Science. 1985 Mar 22;227(4693):1435-41 PMID: 2983426
  45. Yeast chromosome replication and segregation.
    Microbiol Rev. 1988 Dec;52(4):568-601 PMID: 3070325
  46. The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae.
    Science. 1988 Jul 15;241(4863):317-22 PMID: 3291120
  47. Genetically essential and nonessential alpha-tubulin genes specify functionally interchangeable proteins.
    Mol Cell Biol. 1986 Nov;6(11):3722-33 PMID: 3540600
  48. Isolation of the gene encoding yeast DNA polymerase I.
    Cell. 1985 Nov;43(1):369-77 PMID: 3907855
  49. Repetitive zinc-binding domains in the protein transcription factor IIIA from Xenopus oocytes.
    EMBO J. 1985 Jun;4(6):1609-14 PMID: 4040853
  50. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
    Anal Biochem. 1983 Jul 1;132(1):6-13 PMID: 6312838
  51. Separation of chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresis.
    Nucleic Acids Res. 1984 Jul 25;12(14):5647-64 PMID: 6379602
  52. Determination of probability of plasmid loss per generation.
    Gene. 1984 May;28(2):237-9 PMID: 6735179
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.12.12.5736-5747.1992
Published
1992-12-00
Pages
5736-47
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360513
Subset
IM
Grants
NCI NIH HHS · CA16519 · United States
Databases
GENBANK
M94769
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1128/mcb.12.12.5736-5747.1992 PMC Full Text View journal details
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