Staff Profile
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
| Code | Description | Percentage |
|---|---|---|
| 060199 | Biochemistry And Cell Biology Not Elsewhere Classified | 70 |
| 060599 | Microbiology Not Elsewhere Classified | 20 |
| 030499 | Medicinal And Biomolecular Chemistry Not Elsewhere Classified | 10 |
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
Grants and Funding
Summary
| Number of grants | 10 |
|---|---|
| Total funding | $2,296,755 |
For project grants received where the lead institution is other than the University of Newcastle, details are shown in italics.
Click on a grant title below to expand the full details for that specific grant.
2013 (2 grants)
DVC(R) Research Support for Future Fellow (FT12)$60,000
Funding Body: University of Newcastle
| Project Team | ||
|---|---|---|
| Doctor Ian Grainge | ||
| Scheme | Role | |
| Special Project Grant | Chief Investigator | |
| Total Amount | Funding Start | Funding Finish |
| $60,000 | 2013 | 2016 |
| GNo:G1201101 | ||
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 | ||
| Scheme | Role | |
| Travel Grant | Chief Investigator | |
| Total Amount | Funding Start | Funding Finish |
| $1,015 | 2013 | 2013 |
| 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 | ||
| Scheme | Role | |
| Future Fellowships | Chief Investigator | |
| Total Amount | Funding Start | Funding Finish |
| $709,318 | 2012 | 2016 |
| 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 | ||
| Scheme | Role | |
| Project Grant | Investigator | |
| Total Amount | Funding Start | Funding Finish |
| $311,439 | 2012 | 2014 |
| 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 | ||
| Scheme | Role | |
| Equipment Grant | Investigator | |
| Total Amount | Funding Start | Funding Finish |
| $10,000 | 2012 | 2012 |
| 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 | ||
| Scheme | Role | |
| Project Grant | Chief Investigator | |
| Total Amount | Funding Start | Funding Finish |
| $455,022 | 2011 | 2013 |
| GNo:G1000271 | ||
Chemical Biology$444,961
Funding Body: University of Newcastle
| Project Team | ||
|---|---|---|
| Professor Adam McCluskey, Doctor Warwick Belcher, Doctor Ian Grainge, Professor Christopher Grof, Professor Peter Lewis, Professor Eileen McLaughlin, Doctor Shaun Roman, Conjoint Professor Ray Rose, Doctor Jennette Sakoff, Doctor Nikki Verrills | ||
| Scheme | Role | |
| Priority Research Centre | Investigator | |
| Total Amount | Funding Start | Funding Finish |
| $444,961 | 2011 | 2015 |
| GNo:G1100052 | ||
DNA Replication fork processing and recovery in living Escherichia coli cells$285,000
Funding Body: ARC (Australian Research Council)
| Project Team | ||
|---|---|---|
| Doctor Ian Grainge | ||
| Scheme | Role | |
| Discovery Projects | Chief Investigator | |
| Total Amount | Funding Start | Funding Finish |
| $285,000 | 2011 | 2013 |
| 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 | ||
| Scheme | Role | |
| Equipment Grant | Investigator | |
| Total Amount | Funding Start | Funding Finish |
| $15,000 | 2011 | 2011 |
| GNo:G1100028 | ||
2010 (1 grants)
Chromosome stability in pathogenic bacteria$5,000
Funding Body: University of Newcastle
| Project Team | ||
|---|---|---|
| Doctor Ian Grainge | ||
| Scheme | Role | |
| New Staff Grant | Chief Investigator | |
| Total Amount | Funding Start | Funding Finish |
| $5,000 | 2010 | 2010 |
| GNo:G1000625 | ||
Research Supervision
| Number of current supervisions | 5 |
|---|---|
| Total current UoN PhD EFTSL | 2.5 |
For supervisions undertaken at an institution other that the University of Newcastle, details are shown in italics, and the institution name is listed below the program name.
Current Supervision
| Commenced | Proposed Completion | Program | Supervisor Type | Research Title |
|---|---|---|---|---|
| 2013 | 2017 | PhD (Biological Sciences) | Principal Supervisor | Isolation, Purification and Crystallisation of the Major Proteins Involved in the Later Stages of Prokaryotic Chromosome Segregation and Cell Division |
| 2013 | 2017 | PhD (Biological Sciences) | Principal Supervisor | Replication Fork Stalling and Recovery in Escherichia Coli |
| 2012 | 2016 | PhD (Biological Sciences) | Co-Supervisor | Bacterial Transcription Factors NusB, NusE ATFA and p as Targets for Antibiotic Research |
| 2012 | 2016 | PhD (Biological Sciences) | Principal Supervisor | Development of Genetic Tools for the Quantitative Assessment of Gene Depletion in Pathogenic Bacteria |
| 2010 | 2014 | PhD (Biological Sciences) | Co-Supervisor | Structural Characterisation of Bacterial Protein Factors That Bind RNA Polymerase |
Dr Ian Grainge
| Work Phone | 4921 7238 |
|---|---|
| Fax | 4921 5472 |
| Ian.Grainge@newcastle.edu.au | |
| Position | ARC Future Fellow School of Environmental and Life Sciences Faculty of Science and Information Technology |
| Focus Area | Biological Sciences |
| Office | BG09, Biological Sciences, Callaghan University Drive Callaghan NSW 2308 Australia |
| URL: | www.newcastle.edu.au/profile/ian-grainge |
