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Academic Background

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

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.

Figures

Uetake and Sluder Figure 2

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 1

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.

Rotation Projects

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.

One or more keywords matched the following items that are connected to Sluder, Greenfield
Item TypeName
Academic Article Controls for centrosome reproduction in animal cells: issues and recent observations.
Academic Article CDK1 inactivation regulates anaphase spindle dynamics and cytokinesis in vivo.
Academic Article The apparent linkage between centriole replication and the S phase of the cell cycle.
Academic Article Requirement of Cdk2-cyclin E activity for repeated centrosome reproduction in Xenopus egg extracts.
Academic Article Requirement of a centrosomal activity for cell cycle progression through G1 into S phase.
Academic Article Centrosome reproduction in Xenopus lysates.
Academic Article The coordination of centrosome reproduction with nuclear events of the cell cycle in the sea urchin zygote.
Academic Article De novo formation of centrosomes in vertebrate cells arrested during S phase.
Academic Article Mutant Rac1B expression in Dictyostelium: effects on morphology, growth, endocytosis, development, and the actin cytoskeleton.
Academic Article Cell cycle progression and de novo centriole assembly after centrosomal removal in untransformed human cells.
Academic Article Functional properties of kinetochores in animal cells.
Academic Article Relationship between nuclear DNA synthesis and centrosome reproduction in sea urchin eggs.
Academic Article Cell cycle progression after cleavage failure: mammalian somatic cells do not possess a "tetraploidy checkpoint".
Academic Article The de novo centriole assembly pathway in HeLa cells: cell cycle progression and centriole assembly/maturation.
Academic Article Cyclin E in centrosome duplication and reduplication in sea urchin zygotes.
Academic Article MdmX regulates transformation and chromosomal stability in p53-deficient cells.
Academic Article Abrogation of the postmitotic checkpoint contributes to polyploidization in human papillomavirus E7-expressing cells.
Concept S Phase
Concept Cell Division
Concept G1 Phase
Academic Article Link between DNA damage and centriole disengagement/reduplication in untransformed human cells.
Academic Article One to only two: a short history of the centrosome and its duplication.
Concept G2 Phase Cell Cycle Checkpoints
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  • Cell Division