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Career Summary

Biography

I am interested in all aspects of how bacteria pass on their genetic information, from DNA replication to chromosome segregation and accurate cell division.

My current research focuses on two main topics. Firstly what happens when the process of DNA replication runs into a blockage and stops- how can the cell recover to restart the vital process of copying its DNA? A large number of homologous recombination proteins have been implicated in the processing of collapsed DNA replication forks and their roles, and the pathways used, will be investigated in living cells.

Secondly, is the study of the FtsK protein, which co-ordinates the processes of cell division, chromosome unlinking and chromosome segregation in bacteria. Each of these processes has to be completed in a timely manner to allow the cell to divide to produce offspring with a full genetic content. The DNA translocase protein, FtsK, is a key protein in each of these processes, and could additionally act as a cell division checkpoint.

Post-doctoral work:

Department of Biochemistry, University of Oxford (2005-2009)

Cancer Research UK (2000-2004)

University of Texas at Austin (1997-2000)

PhD:

University of Oxford, UK (1994-1997)

Qualifications

  • PhD, University of Oxford - UK, 17/07/1999
  • Bachelor of Arts, University of Cambridge - UK, 24/06/1994
  • Master of Arts, University of Cambridge - UK, 17/10/1998

Research

Research keywords

  • Cell division
  • Chromosome segregation
  • DNA translocation
  • Microbiology
  • Novel antibiotics
  • Site-specific recombination

Research expertise

DNA replication in bacteria: restart of stalled replication forks

The chromosome of E. coli is a circular DNA molecule which is replicated bi-directionally from a single origin (OriC). Multiple copies of the tetracycline operator (tetO) have been placed in the chromosome 16kb to one side of the origin of replication. Expression of a fluorescent tetracycline repressor (TetR-YFP) allows direct visualization of this region in a fluorescence microscope. Using this system, the origin can effectively be followed during duplication and on through the cell cycle. It was found that overexpression of tetR led to cell inviability. The viability of the cells could be recovered by addition of the effector molecule, anhydrous tetracycline (AT) which reduces the binding of TetR to tetO.

Analysis of replication in cells overexpressing TetR showed that the array formed an effective block to replication forks. 2-D gel analysis shows that replication forks can proceed fewer than 500 bp into the tetO array before stalling occurs. Further it is seen that addition of AT leads to a very rapid restart of these stalled forks. Restart of the forks was examined in recA and recB mutant strains and it was found that restart occurred with very similar kinetics to those seen in a wild-type background. This has led us to propose that a stalled replication fork is stable in E. coli for a period of at least 2 hours and that restart does not require recombination. Current work is focusing on what happens when the replisome, stalled at the tetO array block, is disassembled, with a view to understanding replication fork restart pathways and kinetics in vivo. Using this system replication restart can be followed in various mutant backgrounds by examination of fluorescent repressor operators, and by using 2-D gels, to effectively dissect in vivo restart pathways.

FtsK: a fast molecular motor

The multifunctional FtsK protein is involved in cell division and DNA segregation in E. coli. The C-terminal portion of this large protein forms a hexameric ring with the ability to translocate DNA at speeds of ~ 5kb/sec. The FtsK protein is loaded on DNA in a specific orientation by interactions with polarized sequences on the chromosome which ensure that the protein will subsequently move towards the dif site located in the terminus of the chromosome. Once there, FtsK also interacts with the site-specific recombinase XerD to promote recombination between two dif sites. Further, as a result of translocation the two recombining dif sites are brought together in a topologically simple manner so that recombination leads to a simplification of topology, and eventually chromosome unlinking. Using a variety of biochemical techniques the mechanism of directed loading upon DNA, DNA translocation and activation of recombination within a specific synapse topology is being investigated. Using covalently linked multimers of the translocase protion of the protein, hexameric rings can be formed within which mutations can be targeted to specific subunits. This allows more defined analysis of the mechanism of loading and translocation than would otherwise be possible.

Languages

  • English

Fields of Research

CodeDescriptionPercentage
060199Biochemistry And Cell Biology Not Elsewhere Classified70
060599Microbiology Not Elsewhere Classified20
030499Medicinal And Biomolecular Chemistry Not Elsewhere Classified10

Memberships

Body relevant to professional practice.

  • Member - Australian Society for Microbiology

Appointments

Future Fellow
ARC (Australia)
01/12/2012

Administrative

Administrative expertise

Member of Institutional Biosafety Committee

Member of Faculty Research and Research Training Committee

Convenor of Biological Sciences seminar series.

Teaching

Teaching keywords

  • BIOL2010
  • BIOL2230
  • BIOL3100
  • CHEM3550

Teaching expertise

Present course taught:

BIOL2010- Biochemistry

CHEM3550 Medicinal Chemistry

BIOL3100 Microbiology

BIOL2230 Biomolecules

Courses lectured at the University of Oxford, Department of Biochemistry:

Modern Molecular Biology: Methods

DNA: Replication and Recombination

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Publications

For publications that are currently unpublished or in-press, details are shown in italics.

Click on a category title below to expand the list of citations for that specific category.

Chapter (5 outputs)

YearCitationAltmetricsLink
2012Doherty GP, Mettrick KA, Grainge IR, Lewis PJ, 'Imaging fluorescent protein fusions in live bacteria', Methods in Microbiology, Academic Press, Kidlington, Ox 107-126 (2012) [B1]

Co-authors: Peter Lewis

2007Grainge I, Sherratt D, 'Site specific recombination', Topics in Current Genetics: Molecular Genetics of Recombination, Springer, Berlin, Germany 1423-1433 (2007) [B1]
2005Grainge IRF, Jayaram M, 'Introduction to site-specific recombination', The Dynamic Bacterial Genome, Cambridge University Press, Cambridge, United Kingdom 33-82 (2005) [B1]
2005Voziyanov Y, Grainge I, Jayaram M, 'Applications of fungal site-specific recombination as a tool in biotechnology and basic biology', 189-210 (2005) [B1]
2002Jayaram M, Grainge IRF, Tribble GD, 'Site-specific DNA recombination mediated by the Flp protein of Saccharomyces cerevisiae', Mobile DNA II, ASM Press, Washington, DC/USA 192-218 (2002) [B1]
Show 2 more

Journal article (30 outputs)

YearCitationAltmetricsLink
2014Guo P, Grainge I, Zhao Z, Vieweger M, 'Two classes of nucleic acid translocation motors: rotation and revolution without rotation.', Cell Biosci, 4 54 (2014)
2013Shimokawa K, Ishihara K, Grainge I, Sherratt DJ, Vazquez M, 'FtsK-dependent XerCD-dif recombination unlinks replication catenanes in a stepwise manner.', Proc Natl Acad Sci U S A, 110 20906-20911 (2013) [C1]
2013Grainge I, 'Simple topology: FtsK-directed recombination at the dif site', BIOCHEMICAL SOCIETY TRANSACTIONS, 41 595-600 (2013) [C1]
2011Grainge IR, Lesterlin C, Sherratt DJ, 'Activation of XerCD-dif recombination by the FtsK DNA translocase', Nucleic Acids Research, 39 5140-5148 (2011) [C1]
2010Crozat E, Grainge IR, 'FtsK DNA translocase: The fast motor that knows where it's going', ChemBioChem, 11 2232-2243 (2010) [C1]
2010Grainge IR, 'FtsK - a bacterial cell division checkpoint?', Molecular Microbiology, 78 1055-1057 (2010) [C1]
2010Sherratt DJ, Arciszewska LK, Crozat E, Graham JE, Grainge I, 'The Escherichia coli DNA translocase FtsK', BIOCHEMICAL SOCIETY TRANSACTIONS, 38 395-398 (2010) [C1]
2010Crozat E, Meglio A, Allemand J-F, Chivers CE, Howarth M, Venien-Bryan C, et al., 'Separating speed and ability to displace roadblocks during DNA translocation by FtsK', EMBO JOURNAL, 29 1423-1433 (2010) [C1]
2008Grainge I, 'Sporulation: SpoIIIE is the key to cell differentiation', CURRENT BIOLOGY, 18 R871-R872 (2008) [C1]
2008Atanassova N, Grainge I, 'Biochemical Characterization of the Minichromosome Maintenance (MCM) Protein of the Crenarchaeote Aeropyrum pernix and Its Interactions with the Origin Recognition Complex (ORC) Proteins', BIOCHEMISTRY, 47 13362-13370 (2008) [C1]
2008Lowe J, Ellonen A, Allen MD, Atkinson C, Sherratt DJ, Grainge I, 'Molecular mechanism of sequence-directed DNA loading and translocation by FtsK', MOLECULAR CELL, 31 498-509 (2008) [C1]
2007Grainge I, Bregu M, Vazquez M, Sivanathan V, Ip SCY, Sherratt DJ, 'Unlinking chromosome catenanes in vivo by site-specific recombination', EMBO JOURNAL, 26 4228-4238 (2007) [C1]
2007Grainge I, Sherratt DJ, 'Site-specific recombination', Topics in Current Genetics, 17 27-52 (2007)
2006Grainge I, Gaudier M, Schuwirth BS, Westcott SL, Sandall J, Atanassova N, Wigley DB, 'Biochemical analysis of a DNA replication origin in the archaeon Aeropyrum pernix', JOURNAL OF MOLECULAR BIOLOGY, 363 355-369 (2006) [C1]
2006Possoz C, Filipe SR, Grainge I, Sherratt DJ, 'Tracking of controlled Escherichia coli replication fork stalling and restart at repressor-bound DNA in vivo', EMBO JOURNAL, 25 2596-2604 (2006) [C1]
2004Singleton MR, Morales R, Grainge I, Cook N, Isupov MN, Wigley DB, 'Conformational changes induced by nucleotide binding in Cdc6/ORC from Aeropyrum pernix', JOURNAL OF MOLECULAR BIOLOGY, 343 547-557 (2004) [C1]
2003Grainge I, Scaife S, Wigley DB, 'Biochemical analysis of components of the pre-replication complex of Archaeoglobus fulgidus', NUCLEIC ACIDS RESEARCH, 31 4888-4898 (2003) [C1]
2002Grainge I, Pathania S, Vologodskii A, Harshey RM, Jayaram M, 'Symmetric DNA sites are functionally asymmetric within Flp and Cre site-specific DNA recombination synapses', JOURNAL OF MOLECULAR BIOLOGY, 320 515-527 (2002) [C1]
2002Sau AK, Tribble GD, Grainge I, Frohlich RF, Knudsen BR, Jayaram M, 'Biochemical and kinetic analysis of the RNase active sites of the integrase/tyrosine family site-specific recombinases. (vol 276, pg 46612, 2001)', JOURNAL OF BIOLOGICAL CHEMISTRY, 277 6758-6758 (2002)
2002Sau AK, DeVue Tribble G, Grainge I, Frøhlich RF, Knudsen BR, Jayaram M, 'Erratum: Biochemical and kinetic analysis of the RNase active sites of the integrase/tyrosine family site-specific recombinases (Journal of Biological Chemistry (2001) 276 (46612-46623))', Journal of Biological Chemistry, 277 6758-6758 (2002)
2001Frohlich RF, Hansen SG, Lisby M, Grainge I, Westergaard O, Jayaram M, Knudsen BR, 'Inhibition of Flp recombinase by the topoisomerase I-targeting drugs, camptothecin and NSC-314622', JOURNAL OF BIOLOGICAL CHEMISTRY, 276 6993-6997 (2001) [C1]
2001Grainge I, Lee J, Xu CJ, Jayaram M, 'DNA recombination and RNA cleavage activities of the Flp protein: Roles of two histidine residues in the orientation and activation of the nucleophile for strand cleavage', JOURNAL OF MOLECULAR BIOLOGY, 314 717-733 (2001) [C1]
2001Sau AK, Tribble GD, Grainge IRF, Frohlich RF, Knudsen BR, Jayaram M, 'Biochemical and kinetic analysis of the RNase active sites of the integrase/tyrosine family site-specific recombinases', Journal of Biological Chemistry, 276 4661-4662 (2001) [C1]
2001San AK, Tribble GD, Grainge I, Frohlich RF, Knudsen BR, Jayaram M, 'Biochemical and kinetic analysis of the RNase active sites of the integrase/tyrosine family site-specific DNA recombinases', JOURNAL OF BIOLOGICAL CHEMISTRY, 276 46612-46623 (2001)
2000Grainge I, Buck D, Jayaram M, 'Geometry of site alignment during int family recombination: Antiparallel synapsis by the Flp recombinase', JOURNAL OF MOLECULAR BIOLOGY, 298 749-764 (2000) [C1]
1999Grainge I, Sherratt DJ, 'Xer site-specific recombination - DNA strand rejoining by recombinase XerC', JOURNAL OF BIOLOGICAL CHEMISTRY, 274 6763-6769 (1999) [C1]
1999Lee J, Jayaram M, Grainge I, 'Wild-type Flp recombinase cleaves DNA in trans', EMBO JOURNAL, 18 784-791 (1999) [C1]
1998Xu CJ, Grainge I, Lee J, Harshey RM, Jayaram M, 'Unveiling two distinct ribonuclease activities and a topoisomerase activity in a site-specific DNA recombinase', MOLECULAR CELL, 1 729-739 (1998) [C1]
1997Arciszewska L, Grainge IRF, Sherratt DJ, 'Action of site-specific recombinases XerC and XerD on tethered Holliday junctions', The EMBO Journal, 16 3731-3743 (1997) [C1]
1995Arciszewska L, Grainge IRF, Sherratt DJ, 'EFFECTS OF HOLLIDAY JUNCTION POSITION ON XER-MEDIATED RECOMBINATION IN-VITRO', The EMBO Journal, 14 2651-2660 (1995) [C1]
Show 27 more

Review (1 outputs)

YearCitationAltmetricsLink
1999Grainge I, Jayaram M, 'The integrase family of recombinases: organization and function of the active site', MOLECULAR MICROBIOLOGY (1999) [D1]
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Grants and Funding

Summary

Number of grants13
Total funding$2,321,884

- Indicates that the researcher may be seeking students for this project.

Click on a grant title below to expand the full details for that specific grant.

2014 (1 grants)

DNA Replication fork stability, collapse and processing in living Escherichia coli cells$10,000
Funding Body: University of Newcastle

Project Team
Doctor Ian Grainge
SchemeRole
Near Miss GrantChief Investigator
Total AmountFunding StartFunding Finish
$10,00020142014
GNo:G1301382

2013 (4 grants)

DVC(R) Research Support for Future Fellow (FT12)$60,000
Funding Body: University of Newcastle

Project Team
Doctor Ian Grainge
SchemeRole
Special Project GrantChief Investigator
Total AmountFunding StartFunding Finish
$60,00020132016
GNo:G1201101

Development of new genetic tools for protein knockouts in pathogenic bacteria$13,129
Funding Body: University of Newcastle - Faculty of Science & IT

Project Team
Doctor Ian Grainge
SchemeRole
Strategic Small GrantChief Investigator
Total AmountFunding StartFunding Finish
$13,12920132013
GNo:G1401062

Faculty PVC Conference Assistance Grant 2013$2,000
Funding Body: University of Newcastle - Faculty of Science & IT

Project Team
Doctor Ian Grainge
SchemeRole
PVC Conference Assistance GrantChief Investigator
Total AmountFunding StartFunding Finish
$2,00020132013
GNo:G1401158

Characterisation of the interaction between the essential bacterial transcription factor NusA and RNA Polymerase$1,015
Funding Body: Australian Synchrotron

Project Team
Doctor Ian Grainge
SchemeRole
Travel GrantChief Investigator
Total AmountFunding StartFunding Finish
$1,01520132013
GNo:G1301024

2012 (3 grants)

Characterization of a powerful molecular motor, the FtsK DNA translocase$709,318
Funding Body: ARC (Australian Research Council)

Project Team
Doctor Ian Grainge
SchemeRole
Future FellowshipsChief Investigator
Total AmountFunding StartFunding Finish
$709,31820122016
GNo:G1101070

Targeting nucleic acid synthesis and cell division in gram-negative bacterial pathogens$311,439
Funding Body: NHMRC (National Health & Medical Research Council)

Project Team
Professor Nicholas Dixon, Associate Professor Elizabeth Harry, Professor Peter Lewis, Associate Professor Aaron Oakley, Doctor Ian Grainge
SchemeRole
Project GrantInvestigator
Total AmountFunding StartFunding Finish
$311,43920122014
GNo:G1101133

Ultrasonic Homogenizer System and -80 ºC Freezers for chemical and biological sample storage$10,000
Funding Body: NHMRC (National Health & Medical Research Council)

Project Team
Professor Adam McCluskey, Professor Eileen McLaughlin, Professor Peter Lewis, Ms Belinda Nixon, Doctor Shaun Roman, Doctor Jennette Sakoff, Doctor Ian Grainge
SchemeRole
Equipment GrantInvestigator
Total AmountFunding StartFunding Finish
$10,00020122012
GNo:G1100986

2011 (4 grants)

Molecular characterization of the role of FtsK in chromosome unlinking and segregation.$455,022
Funding Body: NHMRC (National Health & Medical Research Council)

Project Team
Doctor Ian Grainge
SchemeRole
Project GrantChief Investigator
Total AmountFunding StartFunding Finish
$455,02220112013
GNo:G1000271

Chemical Biology$444,961
Funding Body: University of Newcastle

DNA Replication fork processing and recovery in living Escherichia coli cells$285,000
Funding Body: ARC (Australian Research Council)

Project Team
Doctor Ian Grainge
SchemeRole
Discovery ProjectsChief Investigator
Total AmountFunding StartFunding Finish
$285,00020112013
GNo:G1000148

Eppendorf mastercycler pro with thermomixer comfort and 5430R centrifuge$15,000
Funding Body: NHMRC (National Health & Medical Research Council)

Project Team
Professor Eileen McLaughlin, Professor Peter Lewis, Professor Adam McCluskey, Conjoint Professor Keith Jones, Associate Professor Brett Nixon, Doctor Shaun Roman, Doctor Jennette Sakoff, Doctor Ian Grainge, Doctor Janet Holt, Doctor Xiao Yang
SchemeRole
Equipment GrantInvestigator
Total AmountFunding StartFunding Finish
$15,00020112011
GNo:G1100028

2010 (1 grants)

Chromosome stability in pathogenic bacteria$5,000
Funding Body: University of Newcastle

Project Team
Doctor Ian Grainge
SchemeRole
New Staff GrantChief Investigator
Total AmountFunding StartFunding Finish
$5,00020102010
GNo:G1000625
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Research Supervision

Number of current supervisions6
Total current UoN PhD EFTSL3.1

For supervisions undertaken at an institution other that the University of Newcastle, the institution name is listed below the program name.

Current Supervision

CommencedProposed
Completion
ProgramSupervisor TypeResearch Title
20142018PhD (Biological Sciences)Principal SupervisorFrequency of Chromosome Dimers Resulting From Replication Fork Processing Following Blockage or Collapse
20132017PhD (Biological Sciences)Principal SupervisorUnderstanding Chromosome Dimer Resolution Systems of a Pathogenic Bacteria at a Molecular Level
20132017PhD (Biological Sciences)Principal SupervisorDNA Replication Fork Processing and Recovery in Escherichia Coli
20122016PhD (Biological Sciences)Co-SupervisorBacterial Transcription Factors NusB, NusE ATFA and p as Targets for Antibiotic Research
20122016PhD (Biological Sciences)Principal SupervisorDevelopment of Genetic Tools for the Quantitative Assessment of Gene Depletion in Pathogenic Bacteria
20102014PhD (Biological Sciences)Co-SupervisorIncreased Understanding of the Molecular Interactions Involved in Bacterial Transcription and Recombination
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Dr Ian Grainge

Work Phone4921 7238
Fax4921 5472
Email
PositionARC Future Fellow
School of Environmental and Life Sciences
Faculty of Science and Information Technology
The University of Newcastle, Australia
Focus AreaBiological Sciences
Office
BG09,
Biological Sciences,
Callaghan
University Drive
Callaghan NSW 2308
Australia
URL:www.newcastle.edu.au/profile/ian-grainge