Postdoctoral Position Available

Lab Page Link

Greenfield Sluder, Ph.D.

Academic Role: Professor

Faculty Appointment(s) In:
   Cell Biology

Other Affiliation(s):
   Cell Dynamics Group
   Interdisciplinary Graduate Program

Mitosis

Our research is centered on the mechanisms that control various aspects of cell division. Applying microscopic and biophysical methods, we seek to elucidate the functional properties of control mechanisms as they operate in the living cell. Our results establish the basis for the integration of cell function with advances in the molecular biology of regulatory pathways. We use echinoderm zygotes, frog egg extracts, and cultured cells as model systems.

One of the major projects in the laboratory seeks to elucidate the controls that ensure that the interphase centrosome reproduces, or doubles, only once in each cell cycle in proper coordination with nuclear events.  We have recently shown that centrosome reproduction is coordinated with nuclear events by activities that function during S phase of the cell cycle.

We have also developed the first Xenopus egg extract system that supports repeated rounds of centrosome reproduction in vitro.  Using this system we have shown that the activity of the cyclin dependent kinase 2- cyclin E complex (Cdk2-cyclin E) is required for multiple rounds of centrosome duplication.

For Recent Images and Data, Please Visit our Lab Webpage:

http://sluderlab.googlepages.com

Figures

Figure 1:   Mammalian cells. (A and B) Overlaid phase and fluorescence images showing BrdU incorporation in mononucleate and binucleate cells. (A) Cells were previously treated with 0.5 µM cytochalasin D and cultured on bare glass. Mononucleate cells have incorporated BrdU, whereas the binucleates have not. (B) Cells treated with 0.5 µM cytochalasin D and cultured on fibronectin-coated glass. Both the mononucleate and binucleate cells have incorporated BrdU. (C) Cells previously treated with 0.5 µM cytochalasin D and cultured on fibronectin-coated glass (images taken from Video 4, available at http://www.jcb.org/cgi/content/full/jcb.200403014/DC1). Frames from a video sequence of two binucleate cells in the same field progressing through mitosis. The first to enter mitosis (top row) divides into two, whereas the second (bottom row) divides into three. Phase-contrast microscopy. Times are in h:min after cytochalasin D removal. Bars, 50 µm.

Figure 2:  Repeated rounds of centrosome duplication in an aphidicolin-treated Xenopus egg extract.  Frames from a time-lapse video sequence, showing the increase in aster number over time in a microscope field.  The decrease in aster number in panel d is due to the migration of asters from the plane of focus and field of view.  Minutes after addition of sperm nuclei are seen in the lower right corner of each frame.  Polarization optics. 10 microns per scale division.

In other studies we have characterized the cell cycle checkpoint controls for the metaphase-anaphase transition. These checkpoint pathways serve to ensure the equal distribution of chromosomes during cell division. We have demonstrated that signal transducing molecules in the kinetochore monitor chromosome attachment to the spindle and that even one unattached kinetochore will block the metaphase-anaphase transition.

Using GFP-cyclin B and confocal microscopy we are also investigating how the checkpoint pathway that monitors the completion of DNA synthesis controls nuclear envelope breakdown and entry of the cell into mitosis.

Recent Publications

Sluder G.. 2005.Two-way traffic: Centrosomes and the cell cycle. Nat Rev Mol Cell Biol. 2005 Sep;6(9):743-8.

Ehrhardt A. and Sluder G.. 2005. Spindle Pole Fragmentation Due to Proteasome Inhibition. J. Cell. Physiol.  2005 Sep;204(3):808-18.

La Terra S., English C.N.,  Hergert P., McEwen B.F.,  Sluder G., and A. Khodjakov. 2005. The de novo centriole assembly pathway in HeLa cells: cell cycle progression and centriole assembly/maturation.  J. Cell Biol. 149:317-330.

Uetake Y.  and G. Sluder 2005. Cell cycle progression after cleavage failure: Mammalian somatic cells do not possess a “tetraploidy checkpoint” J. Cell Biol. 165:609-15.

Murata-Hori M., Sluder G., and Wang Y.L.. 2004. Regulation of cell cycle by the anaphase spindle midzone. BMC Cell Biol. 5:49.

Sluder G. , Hinchcliffe E.H., and Rieder C.L. (2004). The progression and regulation of mitotic events. In G. Stein and A. Pardee (Eds.). Cell cycle and growth control: Biological Regulation and cancer. John Wiley and Sons, Hoboken NJ, In press.

Sluder G., Nordberg J.J., Miller, F.J., and Hinchcliffe E.H. (2004). A sealed preparation for long term observations of cultured cells. In Live cell imaging: A laboratory manual (ed. D.L. Spector and R.D. Goldman). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York. (In press).

Sluder G.  (2004). Centrosome duplication and its regulation in the higher animal cell.  Chapter 9 In: EA Nigg (Ed.)  Centrosomes in Development and Disease, Wiley-VCH, Weinheim, pp167-189.

Sluder G.  and Nordberg J.J. (2004) The good, the bad and the ugly: the practical consequences of centrosome amplification. Curr Opin Cell Biol. Feb;16(1):49-54.

Hinchcliffe E.H. and Sluder G. (2003). "Do not (mis-)adjust your set": maintaining specimen detail in the video microscope.  Methods Cell Biol. 2003;72:65-85.

Sluder G. and Nordberg J.J. (2003). Microscope basics.  Methods Cell Biol. 2003;72:1-10.

Sluder G. and Wolf D.E.  (Eds.) (2003). Digital Microscopy Methods Cell Biol . Vol. 72 Academic Press, San Diego.

Hinchcliffe E. H., and G. Sluder . (2002). Two for two: Cdk2 and its Role in Centrosome Doubling. Oncogene 21:6154-6160.

Khodjakov A., Rieder C.L., Sluder G. , Cassels G., Sibon O., and Wang C. (2002). De novo formation of centrosomes in vertebrate cells arrested during S phase. J. Cell Biol. 158: 1171-1181.

Hinchliffe E.H. and G. Sluder. (2001). Centrosome duplication: Three kinases come up a winner!. Curr. Biol. 11 (17): R698-R701 SEP 4 2001

Hinchliffe E.H. and G. Sluder . (2001). It takes two to tango: Understanding how centrosome duplication is regulated throughout the cell cycle.  Genes and Development 15:1167-1181.

Hinchcliffe E.H., and G. Sluder , (2001). Preparation of Xenopus egg extracts for the study of centrosome duplication.  Methods in Cell Biology . R.E. Palazzo ed. Academic Press, San Diego. Vol. 67: 269-287 .

Piel, M., J. Nordberg, U. Euteneuer, and M. Bornens. (2001). Centrosome-dependent exit of cytokinesis in animal cells. Science 291: 1550-1553.

Hinchcliffe, E.H., F.J. Miller, M. Cham, A. Khodjakov and G. Sluder . (2001). Requirement of a centrosomal activity for cell cycle progession through G1 into S phase. Science 291:1547-1550.

Sluder, G. and D. McCollum (2000). The mad ways of meiosis. Science 289:254-255.

Hinchcliffe, E. H., E. A. Thompson, F. J. Miller, J. Yang, and G. Sluder (1999). Nucleo-cytoplasmic interactions in the control of nuclear envelope breakdown and entry into mitosis in the sea urchin zygote. J. Cell Sci. 112:1139-1148.

Hinchcliffe, E. H., C. Li, E. A. Thompson, J. L. Maller, and G. Sluder (1999). Requirement of Cdk2 - Cyclin E Activity for Repeated Centrosome Reproduction in Xenopus Egg Extracts. Science. 283:851-854.

Potential Rotation Projects

We favor having students work independently on defined projects that have promise of completion during the study period.

Project #1: Use a frog egg extract that supports multiple rounds of centrosome duplication in vitro to investigate the role of various high interest kinases in the control of centrosome duplication. Also, we can use these extracts to further investigate how various other proteins interact with the centrosome to influence its duplication.

Hinchcliffe, E.H., C. Li, E.A. Thompson, J.L. Maller and G. Sluder. 1999. Requirement of Cdk2-cyclin E activity for repeated centrosome reproduction in Xenopus egg extract. Science 283:851-854.

Project #2: Use microsurgery and immunofluorescence on BSC-1 cells to further investigate how centrioles influence the interphase progression of the cell cycle.

Hinchcliffe, E.H., F.J. Miller, M. Cham, A. Khodjakov and G. Sluder. 2001. Requirement of a centrosomal activity for cell cycle progression through G1 into S phase. Science 291: 1547-1550.

Laboratory Personnel

Anna M. Krzywicka-Racka, Post-Doctoral Fellow

Joshua J. Nordberg, Graduate Student

Yumi Uetake, Post-Doctoral Fellow

Shawn A. Galdeen, Post-Doctoral Fellow

Academic Background

Kip Sluder received his A.B. from Middlebury College in 1968 and his Ph.D. from the University of Pennsylvania in 1976. After an American Cancer Society post-doctoral fellowship with Dan Mazia at the University of California, Berkeley (1977 to 1980), he joined the faculty of the Worcester Foundation for Experimental Biology in 1981. Since 1990, he has served as co-director of the Analytical and Quantitative Light Microscopy Course at the Marine Biological Laboratory, in Woods Hole Massachusetts. In 1997 Kip joined the Cell Biology Department at the University of Massachusetts Medical School as a Professor.

For more information, please visit our lab webpage:

http://sluderlab.googlepages.com/

Office: Biotech IV, Suite 324
Phone: 508-856-8651
E-mail: Greenfield.Sluder@umassmed.edu
Keywords: Biophysics, Cell Biology, Cell Cycle

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Postdoctoral Position Available A postdoctoral position is available to study in this laboratory. Contact Dr. Sluder for additional details.