Cell cycle

New Publication in Cell! The Phosphoregulation of Mitosis

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We are incredibly excited to announce that our SnapShot is out today in Cell!
This snapshot of mitosis collates hundreds of phosphorylation events and directly links them with their regulatory kinases and counterbalancing phosphatases, in both time and space, in a highly innovative ‘circtanglar’ cell layout. More importantly, the static PDF version is accompanied by an interactive website that enables users to access direct links to PubMed, UniProt, and Aquaria 3D protein structures for each and every phosphorylation event shown. The pop-up boxes also contain over 100 additional phosphorylation sites on dozens of proteins essential for mitosis. You can access the interactive web version here:  http://www.cell.com/cell/enhanced/odonoghue2
Even better news is that until August 04, 2017 the PDF version of the SnapShot is freely accessible for everyone at the following link https://authors.elsevier.com/a/1VDWh_278yyILK
A big thank-you to Jenny, Sam, Marcos and Sean for helping me put together what I hope will be an amazing resource for anyone interested in how cells divide and phosphorylation in general.

8th Garvan Signalling Symposium – Registration Now Open !!!

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GarvanSignalling20168th Garvan Signalling Symposium

 Date: Monday 31st of October and Tuesday 1st of November 2016
Venue: The Garvan Institute of Medical Research, Sydney
Registration – Abstract submission to 15th of September 2016
 The Garvan International Signalling Symposium is a premier meeting focused on the mechanisms of signal transduction. It began as a small meeting organised by Prof. Roger Daly around the visit of Prof. Axel Ullrich to the Garvan Institute in 2001. Since then, ‘Signalling Meetings’ have been held every 2-3 years and the meeting has grown into one of the Garvan Institute’s premiere scientific events. This boutique symposium highlights cutting-edge developments from the Asia-Pacific region and the rest of the world.
The Garvan International Signalling Symposium is a premier meeting focused on the mechanisms of signal transduction. It began as a small meeting organised by Prof. Roger Daly around the visit of Prof. Axel Ullrich to the Garvan Institute in 2001. Since then, ‘Signalling Meetings’ have been held every 2-3 years and the meeting has grown into one of the Garvan Institute’s premiere scientific events. This boutique symposium highlights cutting-edge developments from the Asia-Pacific region and the rest of the world.

We welcome scientists at all levels, including students, post-docs, research staff and senior lab heads. The intimate nature of the meeting and enjoyable social functions promotes a collegial atmosphere and excellent networking opportunities. A poster session will be held on the Monday afternoon with generous prizes. Slots have been reserved for short (15 minutes) talks to be selected from submitted abstracts.

The meeting is held at the Garvan Institute in the glamorous Darlinghurst region of Sydney, close to the city, Oxford Street, King’s Cross and the harbour.

This years exciting program features state-of-the-art technologies to investigate a wide range of diseases including cancer, immunology, neuroscience and metabolic disorders.  Special sessions focus on in vivo/intravital signalling, proteomics, control of gene regulation and the structural basis of signalling.

Click Here for more information and to register

 

Speakers

Keynote Speakers

Klaus M Hahn

Klaus HahnThe University of North Carolina at Chapel Hill, USA

Dr. Hahn’s laboratory develops new means to visualize and control protein activity in vivo, and uses them to study the role of signaling dynamics in immune cell decision making. His laboratory has produced broadly applicable approaches to fluorescent biosensors that report conformational changes of endogenous proteins, fluorescent dyes to visualize protein activity in vivo, and protein analogs that can be controlled by light or small molecules. Current biological studies focus on phagocytosis, platelet production and metastasis.

Dr. Hahn studied at the University of Pennsylvania and the University of Virginia, where he received his Ph.D in Organic Chemistry. He was a postdoc at the Center for Fluorescence Research at Carnegie Mellon University, became an Associate Professor of Cell Biology at the Scripps Research Institute, and then moved to UNC-Chapel Hill, where he is the Thurman Distinguished Professor of Pharmacology and Director of the UNC-Olympus Imaging Center. Dr. Hahn is a recipient of an NIH Transformative Grant, the NIH’s James Shannon Director’s Award, and is a fellow of the AAAS. His lab’s work on biosensors was named one of the “10 Breakthroughs of the Decade” by Nature Reviews Molecular Cell Biology.

http://www.hahnlab.com/

 

 

Owen J Sansom

Owen J SansomThe Beatson Institute, Scotland

Owen Sansom is interim director of the Cancer Research UK Beatson Institute, Glasgow. Owen gained his PhD in 2001 working on in vivo models of apoptosis in cancer. Since then, he has been instrumental in determining the molecular hallmarks of colorectal cancer (CRC), including showing the roles of the tumour suppressor protein APC and the WNT signalling pathway and the involvement of intestinal stem cells in tumourigenesis. In 2005, Owen established his own laboratory at the Cancer Research UK Beatson Institute, where he became Deputy Director in 2010. The Sansom laboratory uses in vivo models and 3D in vitro models to recapitulate CRC and pancreatic cancer to investigate the molecular mechanisms underpinning tumourigenesis and to identify novel drug targets. In 2007 Owen won the BACR/AstraZeneca Young Scientist Frank Rose Award and in 2012 was elected a Fellow of the Royal Society of Edinburgh and was awarded the Cancer Research UK Future Leaders in Cancer Research prize.

http://www.beatson.gla.ac.uk/invasion-and-metastasis/owen-sansom-colorectal-cancer-and-wnt-signalling.html

 

International Speakers

Eric O'Neill

Eric O’Neill

University of Oxford, UK

Eric O’Neill is a Senior Group Leader and Associate Professor at the CRUK/MRC Oxford Institute for Radiation Oncology. After completing a Ph.D. at the University of Umeå, Sweden he was a post-doc at the University of Oxford. Subsequently, he was awarded a Marie Curie research fellowship and completed a 5-year post-doctoral position investigating oncogenic and tumour suppressor signalling at the CRUK Beatson Institute for Cancer Research in Glasgow. He is a member of the Association for Radiation Research and an examiner for the Royal College of Radiologists. He has also been on the organising committee for several international conferences.

http://www.radiationoncology.ox.ac.uk/research/eric-oneill

 

PR Shepherd

Peter R Shepherd

University of Auckland, New Zealand

Peter Shepherd gained a PhD in Chemistry from Massey University in New Zealand before post doctoral fellowships at Harvard University and  Cambridge University.  Following faculty positions at University College London, he moved back to Auckland University in 2004 where he is currently deputy director at the Maurice Wilkins Centre, a Centre of Research Excellence. His work focusses on understanding how defects in cell signalling pathways contribute to the development of type-2 diabetes and cancer, particularly focussing on the PI 3-kinase and beta-catenin pathways.  His lab also has a strong translational research focus in the form of several ongoing in house drug discovery projects.

https://unidirectory.auckland.ac.nz/profile/peter-shepherd

 
 
 
 

Tilman Brummer

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University of Freiburg, Germany
 

Dr. Tilman Brummer is currently an independent group leader at the Institute of Molecular Medicine and Cell Research (IMMZ) at the Albert-Ludwigs University (ALU) in Freiburg, Germany. He finished his studies in Biology with a diploma at the ALU followed by a PhD thesis on B lymphocyte signalling with Prof. Michael Reth at the Max-Planck-Institute for Immunobiology and Epigenetics. In 2003, he joined the laboratory of Prof. Roger J. Daly at the Garvan Institute of Medical Research in Sydney as a postdoctoral fellow. In 2008, he returned to the newly established Centre of Biological Systems Analysis (ZBSA) at the ALU to establish his independent laboratory funded by the Emmy-Noether program of the German Research Foundation (DFG) before moving to the IMMZ in 2012. He is a principal investigator within the Collaborative Research Centre 850 “Control of Cell Motility in Morphogenesis, Cancer Invasion and Metastasis”, the “Spemann Graduate School of Biology and Medicine” and the “Centre for Biological Signalling Studies” BIOSS. Recently, he has been awarded a prestigious Heisenberg fellowship of the DFG.

The Brummer laboratory is interested in the organisation of intracellular signalling pathways and how their intricate control becomes disturbed in human cancer. The laboratory is particularly interested in understanding the regulation of the BRAF and GAB2 oncoproteins and how they contribute to metastasis and drug resistance in solid tumours and leukaemia.

 

 

Jen Morton

Jennifer Morton

The Beatson Institute, Scotland
 

Jen Morton is a joint leader of the pancreatic cancer research team at the Cancer Research UK Beatson Institute.  Her research focuses mainly on: (a) Investigating the importance of mutations found in human pancreatic tumours using mouse models, (b) Profiling different genetic subsets of pancreatic cancer to better understand the disease and identify specific targets for therapy, and (c) performing preclinical trials of targeted therapies in clinically and genetically relevant pancreatic cancer mouse models.

 

Pat Caswell

Pat Caswell

Wellcome Trust Centre for Cell-Matrix Research, UK

Patrick is based within the Wellcome Trust Centre for Cell Matrix Research at the University of Manchester. Patrick studied Biochemistry at the University of Nottingham, before undertaking a PhD at the University of Leicester and a postdoc in Jim Norman’s lab at the Beatson Institute for Cancer Research. Patrick set up his lab in 2010, focussing on how cell-matrix interactions through integrins generate signals that control key cellular processes such as cell migration, differentiation and survival. The lab is specifically interested in vesicular trafficking, and has recently shown that endocytic trafficking of integrins and co-cargo receptors controls the spatial activation of RhoGTPases to modulate the actin cytoskeleton in invasive cancer cells.

http://www.wellcome-matrix.org/research_groups/pat-caswell.html

 

Vinay Tergaonkar

Vinay Tergaonker

Institute of Molecular and Cell Biology, Singapore
 

Vinay Tergaonkar obtained his Ph.D. (2001), from National Center for Biological Sciences, Bangalore. During his graduate studies he was awarded an international cancer society (UICC) fellowship for collaborative research at Tufts University, Boston, USA. He has been a fellow (2001-2004) and a special fellow (2004-present) of the Leukemia and Lymphoma Society of America and conducted his postdoctoral studies at the Salk Institute for Biological Studies, La Jolla, California. He joined the Institute of Molecular and Cell Biology (IMCB) in late 2005 as Principal Investigator and became a Senior Principal Investigator in 2010 and Research Director in 2015. He is also a Professor at School of Medicine at National University of Singapore. He has been invited to speak at various international venues and meetings such as the Barossa and Hunter valley meetings in Australia, Genes and Cancer meeting in UK, The Argentine Pharmacological society meeting in Buenos Aires, Aichi and Japanese Cancer Society meetings in Japan and the Keystone Symposia. He serves on 
Editorial Boards of 1) Molecular and Cellular Biology (American Society for Molecular Biology) 2) Biochemical Journal (Portland Press) 
3) Critical Reviews in Oncology/Hematology (Elsevier Press), 4) BMC Research Notes (Biomed Central) and 5) Telomeres and Telomerase. He has received international recognition for his work including the British council development award (2014) and the 2015 Premiers’ fellowship from Government of South Australia.

 

Daniel Schramek

Daniel Schramek

Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, University of Toronto, Canada

Daniel Schramek is a Principal Investigator at the Lunenfeld-Tanenbaum Research Institute, and Assistant Professor in the Department of Molecular Genetics, Faculty of Medicine, University of Toronto. He obtained his BSc and MSc in Molecular biology from the University of Vienna and undertook his master thesis under the supervision of Prof. Roger Daly at the Garvan Institute in Sydney. He undertook his PhD work under the supervision of Prof. Josef Penninger at IMBA in Vienna, followed by postdoctoral studies with Prof. Elaine Fuchs at the Rockefeller University in New York. In 2015, Dr. Schramek was recruited to the Lunenfeld-Tanenbaum Research Institute, where he holds an endowed ‘Kierans & Janigan’ Cancer Research Chair as well as a tier 2 Canadian Research Chair in Functional Cancer Genomics. Dr. Schramek was awarded a prestigious HFSP Career Development Award, the Early Researcher Award from the Ontario Ministry of Research & Innovation as well as a CIHR Foundation grant.

His lab focuses on functional cancer genomics and has generated various in vivo CRISPR-gene editing methodologies to screen for novel tumor suppressors, cancer vulnerabilities as well as therapy resistance genes in various mouse models of human cancers. Through this program, Dr. Schramek specifically aims to identify molecular and cellular mechanisms that regulate epithelial tissue growth in adult homeostasis, cancer and cancer-associated inflammation.

http://www.lunenfeld.ca/researchers/schramek

http://schramek.lunenfeld.ca

 

 

BioEssays Review on Mitotic Phosphatase Specificity now Online

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Great News, our review on how phosphatase specificity is controlled during mitosis has been re-published in BioEssays and is now Online!

This review was originally published in Inside the Cell, which unfortunately has shut down.

But the good news is that it is still Open Access, so that means its free for everyone to read! And is now also indexed in PubMed

During mitotic exit, phosphatases reverse thousands of phosphorylation events in a specific temporal order to ensure that cell division occurs correctly. This review explores how the physicochemical properties of the phosphosite and surrounding amino acids affect interactions with phosphatase/s and help determine the dephosphorylation of individual phosphorylation sites during mitotic exit.

The Full-text download for the Article can be found here [Link]

Citation: Rogers, S. et al. (2016) Mechanisms regulating phosphatase specificity and the removal of individual phosphorylation sites during mitotic exit. BioEssays, 38, S24–S32.

 

Switching off Cancers Diversity

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JCS paper

A defining feature in over 2/3rds of all solid tumours is the continual loss and gain of whole are small parts of chromosomes. This instability, or CIN for short, strongly implicated in tumour initiation, progression, chemoresistance and poor prognosis. CIN is created through failures during mitosis, whereby whole or parts of a chromosome are segregated incorrectly, thereby created daughter cells with unequal chromosome numbers. Consequently, understanding how mitosis is regulated is essential for uncovering the mechanisms allowing CIN to arise and drive cancer. In our recent publication, we discovered the mechanisms controlling the key regulatory pathway critical to ensuring cells exit mitosis correctly. At the centre of this pathway is a gene call MASTL (short for ‘Microtubule Associated Serine/Threonine Kinase-Like’). The primary function of MASTL is to ensure that the cellular breaks (the phosphatase PP2A), is turned off during mitosis so that the accelerator (Cdk1 kinase) can drive the cell into mitosis. Much like a car, having the accelerator and breaks on at the same time is a bad idea, unless you like the smell of burning rubber. To successfully exit mitosis, and to perfectly segregate chromosomes, the cell must take the foot off the accelerator and turn on the breaks. Because MASTL is the central regulator ensuring the breaks are coordinated with the accelerator, it is essential to understand how MASTL is controlled. To this end, we uncovered that MASTL must be rapidly turned off to allow cells to exit mitosis, and this inactivation is carried out by another cellular brake call PP1 phosphatase (Rogers et al, JCS 2016). Now that we have identified and mapped this novel mitotic exit switch, we hope to be able to shed new light on how CIN drives the initiation and evolution cancer. We believe that with further study we will be able to better predict patient response to chemotherapy, and also identify new ways to ‘switch off’ highly unstable tumours, thereby improving treatment for patients that currently have a very poor prognosis.

Image of Interphase HeLa cell stained for Actin (red), DNA (blue) and the co-localisation of MASTL and PP1 by Proximity Ligation Assay (PLA; green).
Credit: Sam Rogers and Cell Division Lab

 

New Publication: Clinical Overview of MDM2/X-Targeted Therapies

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Great news, we have a new Mini-Review published in Frontiers Oncology entitled “Clinical Overview of MDM2/X-Targeted Therapies“, which is apart of the Research Topic Human tumor-derived p53 mutants: a growing family of oncoproteins

Here is a little snippet from the Abstract to wet your appetite!

MDM2 and MDMX are the primary negative regulators of p53, which under normal conditions maintain low intracellular levels of p53 by targeting it to the proteasome for rapid degradation and inhibiting its transcriptional activity. Both MDM2 and MDMX function as powerful oncogenes and are commonly over-expressed in some cancers, including sarcoma (~20%) and breast cancer (~15%).

In this overview, we will review the current MDM2- and MDMX-targeted therapies in development, focusing particularly on compounds that have entered into early phase clinical trials. We will highlight the challenges pertaining to predictive biomarkers for and toxicities associated with these compounds, as well as identify potential combinatorial strategies to enhance its anti-cancer efficacy.

The article is Open Access, which means its free for everyone and anyone to read and download!

You can view and download it directly here [Link]

New Review Article Published!! “Mechanisms Regulating Phosphatase Specificity During Mitotic Exit”

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icl31035-toc-0001

Great News, we have a new review article that has just been published online today in Inside the Cell!
Its Open Access, so that means its free for everyone to read!

During mitotic exit, phosphatases reverse thousands of phosphorylation events in a specific temporal order to ensure that cell division occurs correctly. This review explores how the physicochemical properties of the phosphosite and surrounding amino acids affect interactions with phosphatase/s and help determine the dephosphorylation of individual phosphorylation sites during mitotic exit.

The Full Reference and link for the Article can be found below:
Samuel Rogers, Rachael McCloy, D Neil Watkins and Andrew Burgess Mechanisms regulating phosphatase specificity and the removal of individual phosphorylation sites during mitotic exit Inside the Cell [Link]

Position available: 2016 Honours Student Project in our Lab

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Great news we are currently looking for a new honours student for 2016.

The title of the project is “Developing novel biosensors to monitor DNA damage in cancer cells”.

Its a very exciting new project incorporating cutting edge microscopy and fluorescent biosensors.

If you think you have what it takes and are interested please feel free contact myself, or UNSW SoMS.
For more information on the UNSW honours program please visit: http://medicalsciences.med.unsw.edu.au/students/soms-honours/

Below is an example of the images that will be created during the project.

Using ImageJ to Measure Cell Fluorescence

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Image J can be downloaded for free from here .
This guide can also be downloaded as a complete PDF here: Measuring Cell Fluorescence using ImageJ

Here is a very simple guide for determining the level of  fluorescence in a given region (e.g nucleus)

  1. Select the cell of interest using any of the drawing/selection tools (i.e. rectangle, circle, polygon or freeform)
  2. From the Analyze menu select “set measurements”. Make sure you have AREA, INTEGRATED DENSITY and MEAN GRAY VALUE selected (the rest can be ignored).
  3. Now select “Measure” from the analyze menu or hit cmd+m (apple). You should now see a popup box with a stack of values for that first cell.
  4. Now go and select a region next to your cell that has no fluroence, this will be your background.
    NB: the size is not important. If you want to be super accurate here take 3+ selections from around the cell.
  5. Repeat this step for the other cells in the field of view that you want to measure.
  6. Once you have finished, select all the data in the Results window, and copy (cmd+c) and paste (cmd+v) into a new excel worksheet (or similar program)
  7. Use this formula to calculate the corrected total cell fluorescence (CTCF).
    NB: You can use excel to perform this calculation for you.
    CTCF = Integrated Density – (Area of selected cell  X Mean fluorescence of background readings)

     
  8. Make a graph and your done. Notice that in this example that the rounded up mitotic cell appears to have a much higher level of staining, but this is actually due to its smaller size, which concentrates the staining in a smaller space. So if you just used the raw integrated density you would have data suggesting that the flattened cell has less staining then the rounded up one, when in reality they have a similar level of fluorescence.

How to Cite this if you wold like to:

We have used this method in these papers:

McCloy, R. A., Rogers, S., Caldon, C. E., Lorca, T., Castro, A., and Burgess, A. (2014) Partial inhibition of Cdk1 in G 2 phase overrides the SAC and decouples mitotic events. Cell Cycle 13, 1400–1412 [Link]

Burgess A, Vigneron S, Brioudes E, Labbé J-C, Lorca T & Castro A (2010) Loss of human Greatwall results in G2 arrest and multiple mitotic defects due to deregulation of the cyclin B-Cdc2/PP2A balance. Proc Natl Acad Sci USA 107: 12564–12569

But you can also find a similar method published here:

Gavet O & Pines J (2010) Progressive activation of CyclinB1-Cdk1 coordinates entry to mitosis. Dev Cell 18: 533-543

And here:

Potapova TA, Sivakumar S, Flynn JN, Li R & Gorbsky GJ (2011) Mitotic progression becomes irreversible in prometaphase and collapses when Wee1 and Cdc25 are inhibited. Mol Biol Cell 22: 1191–1206

And my apologies to any others that I have not mentioned.

Congratulation to Sam our PhD student on his 1st Author Publication!

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Great news, we recently published a collaborative paper in the journal Cell Division with the Lab of Dr Liz Caldon here at the Garvan.
The title of the paper is “Cyclin E2 is the predominant E-cyclin associated with NPAT in breast cancer cells”, and you can find it online here

It also marked the inaugural 1st Author publication for our PhD student Sam!

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