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WO2006108179A3 - Two component dna pol iii replicases with modified beta-subunit binding motifs, and uses thereof - Google Patents

Two component dna pol iii replicases with modified beta-subunit binding motifs, and uses thereof Download PDF

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Publication number
WO2006108179A3
WO2006108179A3 PCT/US2006/013493 US2006013493W WO2006108179A3 WO 2006108179 A3 WO2006108179 A3 WO 2006108179A3 US 2006013493 W US2006013493 W US 2006013493W WO 2006108179 A3 WO2006108179 A3 WO 2006108179A3
Authority
WO
WIPO (PCT)
Prior art keywords
replicases
subunit
component
subunits
modified
Prior art date
Application number
PCT/US2006/013493
Other languages
French (fr)
Other versions
WO2006108179A2 (en
Inventor
Lars-Erik Peters
Original Assignee
Qiagen North American Holdings
Lars-Erik Peters
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US11/101,977 external-priority patent/US8192960B2/en
Application filed by Qiagen North American Holdings, Lars-Erik Peters filed Critical Qiagen North American Holdings
Priority to JP2008505653A priority Critical patent/JP2008535497A/en
Priority to EP06740859A priority patent/EP1874925A2/en
Publication of WO2006108179A2 publication Critical patent/WO2006108179A2/en
Publication of WO2006108179A3 publication Critical patent/WO2006108179A3/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1241—Nucleotidyltransferases (2.7.7)
    • C12N9/1252—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The invention provides modified DNA polymerase III replicases, and methods of using the same for various nucleic replication applications. The modified replicases comprise two components, particularly a polymerase component containing an α subunit, and a sliding clamp component containing a β subunit. The two component polymerases lack a clamp loader. The α subunits of the modified replicases have one or more mutations in one or more of their β-subunit binding sites. Consequently, the mutated α subunits have increased affinity for β-subunits, and the two component replicases exhibit increased processivity.
PCT/US2006/013493 2005-04-07 2006-04-07 Two component dna pol iii replicases with modified beta-subunit binding motifs, and uses thereof WO2006108179A2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2008505653A JP2008535497A (en) 2005-04-07 2006-04-07 Binary DNAPOLIII replicase containing a modified beta-subunit binding motif and uses thereof
EP06740859A EP1874925A2 (en) 2005-04-07 2006-04-07 Two component dna pol iii replicases with modified beta-subunit binding motifs, and uses thereof

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US11/101,977 2005-04-07
US11/101,977 US8192960B2 (en) 2004-04-07 2005-04-07 One component and two component DNA Pol III replicases and uses thereof
US74100905P 2005-11-29 2005-11-29
US60/741,009 2005-11-29

Publications (2)

Publication Number Publication Date
WO2006108179A2 WO2006108179A2 (en) 2006-10-12
WO2006108179A3 true WO2006108179A3 (en) 2007-02-01

Family

ID=37074130

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2006/013493 WO2006108179A2 (en) 2005-04-07 2006-04-07 Two component dna pol iii replicases with modified beta-subunit binding motifs, and uses thereof

Country Status (3)

Country Link
EP (1) EP1874925A2 (en)
JP (1) JP2008535497A (en)
WO (1) WO2006108179A2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3932935A1 (en) * 2020-06-29 2022-01-05 Centre National de la Recherche Scientifique Antibacterial peptides

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999013060A1 (en) * 1997-09-12 1999-03-18 Enzyco, Inc. Novel thermophilic polymerase iii holoenzyme
WO2006074157A2 (en) * 2005-01-03 2006-07-13 Eppendorf Ag Modified pol iii replicases and uses thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999013060A1 (en) * 1997-09-12 1999-03-18 Enzyco, Inc. Novel thermophilic polymerase iii holoenzyme
WO2006074157A2 (en) * 2005-01-03 2006-07-13 Eppendorf Ag Modified pol iii replicases and uses thereof

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DALRYMPLE BRIAN P ET AL: "A universal protein-protein interaction motif in the eubacterial DNA replication and repair systems", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 98, no. 20, 25 September 2001 (2001-09-25), pages 11627 - 11632, XP002405654, ISSN: 0027-8424 *
DOHRMANN P R ET AL: "A Bipartite Polymerase-Processivity Factor Interaction: Only the Internal beta Binding Site of the alpha Subunit is Required for Processive Replication by the DNA Polymerase III Holoenzyme", JOURNAL OF MOLECULAR BIOLOGY, LONDON, GB, vol. 350, no. 2, 8 July 2005 (2005-07-08), pages 228 - 239, XP004927290, ISSN: 0022-2836 *
ETTINGER A ET AL: "UNIQUE CHARACTERISTICS OF THERMOPHILIC DNA POLYMERASE III HOLOENZYME ALPHA-SUBUNITS FROM THERMUS THERMOPHILUS AND AQUIFEX AOLICUS", FEDERATION OF AMERICAN SOCIETIES FOR EXPERIMENTAL BIOLOGY. ANNUAL MEETING. ABSTRACTS OF PAPERS, XX, XX, vol. 16, no. 4, SUPPL PT01, March 2005 (2005-03-01), pages A847, XP009068491 *
LOPEZ DE SARO FRANCISCO J ET AL: "Competitive processivity-clamp usage by DNA polymerases during DNA replication and repair.", EMBO (EUROPEAN MOLECULAR BIOLOGY ORGANIZATION) JOURNAL, vol. 22, no. 23, 1 December 2003 (2003-12-01), pages 6408 - 6418, XP002405655, ISSN: 0261-4189 *

Also Published As

Publication number Publication date
JP2008535497A (en) 2008-09-04
EP1874925A2 (en) 2008-01-09
WO2006108179A2 (en) 2006-10-12

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