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PMID: 2174103 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Purification of RIP60 and RIP100, mammalian proteins with origin-specific DNA-binding and ATP-dependent DNA helicase activities.

Molecular and cellular biology ·Vol. 10 ·No. 12 ·1990-12-00 ·Pages 6225-35

Dailey L, Caddle MS, Heintz N, Heintz NH

Abstract

Replication of the Chinese hamster dihydrofolate reductase gene (dhfr) initiates near a fragment of stably bent DNA that binds multiple cellular factors. Investigation of protein interactions with the dhfr bent DNA sequences revealed a novel nuclear protein that also binds to domain B of the yeast origin of replication, the autonomously replicating sequence ARS1. The origin-specific DNA-binding activity was purified 9,000-fold from HeLa cell nuclear extract in five chromatographic steps. Protein-DNA cross-linking experiments showed that a 60-kDa polypeptide, which we call RIP60, contained the origin-specific DNA-binding activity. Oligonucleotide displacement assays showed that highly purified fractions of RIP60 also contained an ATP-dependent DNA helicase activity. Covalent radiolabeling with ATP indicated that the DNA helicase activity resided in a 100-kDa polypeptide, RIP100. The cofractionation of an ATP-dependent DNA helicase with an origin-specific DNA-binding activity suggests that RIP60 and RIP100 may be involved in initiation of chromosomal DNA synthesis in mammalian cells.

Related Genes
dhfr
MeSH Terms
Adenosine Triphosphatases/isolation & purification,metabolism Adenosine Triphosphate/metabolism Animals Base Sequence Cell Line Cell Nucleus/metabolism DNA/genetics,metabolism DNA Helicases/isolation & purification,metabolism DNA-Binding Proteins/isolation & purification,metabolism Deoxyribonuclease I HeLa Cells/metabolism Humans Molecular Sequence Data Nucleotide Mapping Protein Binding Restriction Mapping Tetrahydrofolate Dehydrogenase/genetics
Chemicals
DNA-Binding Proteins Adenosine Triphosphate DNA Tetrahydrofolate Dehydrogenase Deoxyribonuclease I Adenosine Triphosphatases DNA Helicases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Dailey L
Laboratory of Molecular Biology, Rockefeller University, New York, New York 10021.
Caddle M S
Heintz N
Heintz N H
References (58)
58 references, click to expand
  1. DNAse footprinting: a simple method for the detection of protein-DNA binding specificity.
    Nucleic Acids Res. 1978 Sep;5(9):3157-70 PMID: 212715
  2. Replication in the amplified dihydrofolate reductase domain in CHO cells may initiate at two distinct sites, one of which is a repetitive sequence element.
    Mol Cell Biol. 1989 Feb;9(2):532-40 PMID: 2710116
  3. DNA bending by negative regulatory proteins: Gal and Lac repressors.
    Genes Dev. 1989 May;3(5):606-11 PMID: 2744457
  4. Structure and associated DNA-helicase activity of a general transcription initiation factor that binds to RNA polymerase II.
    Nature. 1989 Oct 5;341(6241):410-4 PMID: 2477704
  5. Interaction of the H4 autonomously replicating sequence core consensus sequence and its 3'-flanking domain.
    Mol Cell Biol. 1989 Dec;9(12):5464-72 PMID: 2685566
  6. The DNA unwinding element: a novel, cis-acting component that facilitates opening of the Escherichia coli replication origin.
    EMBO J. 1989 Dec 20;8(13):4335-44 PMID: 2556269
  7. Reversion of autonomously replicating sequence mutations in Saccharomyces cerevisiae: creation of a eucaryotic replication origin within procaryotic vector DNA.
    Mol Cell Biol. 1990 Jan;10(1):265-72 PMID: 2403637
  8. Intramolecular DNA triplexes, bent DNA and DNA unwinding elements in the initiation region of an amplified dihydrofolate reductase replicon.
    J Mol Biol. 1990 Jan 5;211(1):19-33 PMID: 2299670
  9. Conserved DNA structures in origins of replication.
    Nucleic Acids Res. 1990 Mar 25;18(6):1609-12 PMID: 2326199
  10. Identification of an origin of bidirectional DNA replication in mammalian chromosomes.
    Cell. 1990 Sep 7;62(5):955-65 PMID: 2393905
  11. RIP60, a mammalian origin-binding protein, enhances DNA bending near the dihydrofolate reductase origin of replication.
    Mol Cell Biol. 1990 Dec;10(12):6236-43 PMID: 2247056
  12. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  13. A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system.
    Nucleic Acids Res. 1981 Jul 10;9(13):3047-60 PMID: 6269071
  14. Methotrexate-resistant Chinese hamster ovary cells have amplified a 135-kilobase-pair region that includes the dihydrofolate reductase gene.
    Proc Natl Acad Sci U S A. 1981 Oct;78(10):6043-7 PMID: 6273843
  15. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis.
    Nucleic Acids Res. 1981 Dec 11;9(23):6505-25 PMID: 6275366
  16. An amplified chromosomal sequence that includes the gene for dihydrofolate reductase initiates replication within specific restriction fragments.
    Proc Natl Acad Sci U S A. 1982 Jul;79(13):4083-7 PMID: 6955792
  17. Cloning of the initiation region of a mammalian chromosomal replicon.
    Nature. 1983 Mar 31-Apr 6;302(5907):439-41 PMID: 6300683
  18. Eukaryotic gene transcription with purified components.
    Methods Enzymol. 1983;101:582-98 PMID: 6888276
  19. Structural requirements for the function of a yeast chromosomal replicator.
    Cell. 1984 May;37(1):299-307 PMID: 6327056
  20. Changes in site specificity of single-strand-specific endonucleases on supercoiled PM2 DNA with temperature and ionic environment.
    Nucleic Acids Res. 1984 Sep 25;12(18):7071-86 PMID: 6091053
  21. Deletion mutations affecting autonomously replicating sequence ARS1 of Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Nov;4(11):2455-66 PMID: 6392851
  22. Polyomavirus enhancer contains multiple redundant sequence elements that activate both DNA replication and gene expression.
    Mol Cell Biol. 1985 Apr;5(4):649-58 PMID: 2985964
  23. Sequence-induced DNA curvature at the bacteriophage lambda origin of replication.
    Nature. 1985 Oct 3-9;317(6036):451-3 PMID: 2995831
  24. Purification of nuclear factor I by DNA recognition site affinity chromatography.
    J Biol Chem. 1986 Jan 25;261(3):1398-408 PMID: 3003068
  25. A mammalian high mobility group protein recognizes any stretch of six A.T base pairs in duplex DNA.
    Proc Natl Acad Sci U S A. 1986 Mar;83(5):1276-80 PMID: 3456586
  26. A putative origin of replication of plasmids derived from Epstein-Barr virus is composed of two cis-acting components.
    Mol Cell Biol. 1985 Aug;5(8):1822-32 PMID: 3018528
  27. DNA helicase activity of SV40 large tumor antigen.
    EMBO J. 1986 Aug;5(8):1939-44 PMID: 3019672
  28. Isolation of the origin of replication associated with the amplified Chinese hamster dihydrofolate reductase domain.
    Proc Natl Acad Sci U S A. 1986 Oct;83(20):7790-4 PMID: 3094015
  29. Bent DNA at a yeast autonomously replicating sequence.
    Nature. 1986 Nov 6-12;324(6092):87-9 PMID: 3785376
  30. Detection, sequence patterns and function of unusual DNA structures.
    Nucleic Acids Res. 1986 Nov 11;14(21):8513-33 PMID: 3786134
  31. The adenine-thymine domain of the simian virus 40 core origin directs DNA bending and coordinately regulates DNA replication.
    Mol Cell Biol. 1986 Dec;6(12):4578-84 PMID: 3025662
  32. A single polypeptide possesses the binding and transcription activities of the adenovirus major late transcription factor.
    Mol Cell Biol. 1986 Dec;6(12):4723-33 PMID: 3796614
  33. Simian virus 40 (SV40) DNA replication: SV40 large T antigen unwinds DNA containing the SV40 origin of replication.
    Proc Natl Acad Sci U S A. 1987 Jan;84(1):16-20 PMID: 3025851
  34. Direct evidence for DNA bending at the lambda replication origin.
    Science. 1987 Apr 24;236(4800):416-22 PMID: 2951850
  35. Helicase action of dnaB protein during replication from the Escherichia coli chromosomal origin in vitro.
    J Biol Chem. 1987 May 15;262(14):6877-85 PMID: 3032979
  36. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor.
    Cell. 1987 Jun 19;49(6):729-39 PMID: 3034432
  37. Yeast regulatory sequences preferentially adopt a non-B conformation in supercoiled DNA.
    Nucleic Acids Res. 1987 Jun 11;15(11):4467-80 PMID: 3035495
  38. DNA sequence requirements for replication of polyomavirus DNA in vivo and in vitro.
    Mol Cell Biol. 1987 Oct;7(10):3694-704 PMID: 2824994
  39. Purification and properties of Drosophila heat shock activator protein.
    Science. 1987 Nov 27;238(4831):1247-53 PMID: 3685975
  40. Two DNA-binding factors recognize specific sequences at silencers, upstream activating sequences, autonomously replicating sequences, and telomeres in Saccharomyces cerevisiae.
    Mol Cell Biol. 1988 Jan;8(1):210-25 PMID: 3275867
  41. Curved helix segments can uniquely orient the topology of supertwisted DNA.
    Cell. 1988 Feb 26;52(4):545-9 PMID: 2830027
  42. The ease of DNA unwinding as a determinant of initiation at yeast replication origins.
    Cell. 1988 Feb 26;52(4):559-67 PMID: 2830028
  43. Escherichia coli DNA helicase I catalyzes a unidirectional and highly processive unwinding reaction.
    J Biol Chem. 1988 Mar 5;263(7):3208-15 PMID: 2830275
  44. Initiation protein induced helix destabilization at the lambda origin: a prepriming step in DNA replication.
    Cell. 1988 Feb 12;52(3):385-95 PMID: 2830983
  45. Transcriptional elements as components of eukaryotic origins of DNA replication.
    Cell. 1988 Mar 11;52(5):635-8 PMID: 3278812
  46. Duplex opening by dnaA protein at novel sequences in initiation of replication at the origin of the E. coli chromosome.
    Cell. 1988 Mar 11;52(5):743-55 PMID: 2830993
  47. Nuclear DNA synthesis in vitro is mediated via stable replication forks assembled in a temporally specific fashion in vivo.
    Mol Cell Biol. 1988 May;8(5):1923-31 PMID: 3386630
  48. Herpes simplex virus DNA replication: the UL9 gene encodes an origin-binding protein.
    Proc Natl Acad Sci U S A. 1988 Aug;85(15):5414-8 PMID: 2840659
  49. Bent DNA functions as a replication enhancer in Saccharomyces cerevisiae.
    Mol Cell Biol. 1988 Jul;8(7):2763-9 PMID: 3043195
  50. Detection of DNA looping due to simultaneous interaction of a DNA-binding protein with two spatially separated binding sites on DNA.
    Proc Natl Acad Sci U S A. 1988 Sep;85(17):6287-91 PMID: 3413096
  51. Transcription factor OTF-1 is functionally identical to the DNA replication factor NF-III.
    Science. 1988 Sep 2;241(4870):1210-3 PMID: 3413485
  52. A model for initiation at origins of DNA replication.
    Cell. 1988 Sep 23;54(7):915-8 PMID: 2843291
  53. Transcriptional activation of initiation of replication from the E. coli chromosomal origin: an RNA-DNA hybrid near oriC.
    Cell. 1988 Oct 7;55(1):113-23 PMID: 2458841
  54. Energetic coupling between DNA bending and base pair opening.
    Proc Natl Acad Sci U S A. 1988 Oct;85(19):7231-5 PMID: 3174629
  55. New beginnings in studies of eukaryotic DNA replication origins.
    Biochim Biophys Acta. 1989 Jan 23;1007(1):1-14 PMID: 2642383
  56. Purification of the human histone H4 gene-specific transcription factors H4TF-1 and H4TF-2.
    Genes Dev. 1988 Dec;2(12B):1700-12 PMID: 3071490
  57. High-resolution mapping of replication fork movement through the amplified dihydrofolate reductase domain in CHO cells by in-gel renaturation analysis.
    Mol Cell Biol. 1989 Feb;9(2):523-31 PMID: 2710115
  58. Mapping replication units in animal cells.
    Cell. 1989 Jun 16;57(6):909-20 PMID: 2544294
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.12.6225-6235.1990
Published
1990-12-00
Pages
6225-35
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362897
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.10.12.6225-6235.1990 PMC Full Text View journal details
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