Daniel P. Kiehart, Professor  

Daniel P. Kiehart

Our intellectual focus is on identifying determinants of cell shape that function during development. Utilizing molecular genetic and reverse genetic approaches in Drosophila, we have shown that conventional nonmuscle myosin is necessary for driving both cell division and post-mitotic cell shape changes for morphogenesis, and cellular locomotions. Currently, we are investigating how myosin elicits cell shape change and how its function is regulated through filament formation, phosphorylation, sub-cellular targeting and small GTP-binding protein function. We are characterizing myosin light chain kinase; a novel myosin VII heavy chain; and additional elements that participate in localizing myosin and transmitting the forces that it produces. We used screens for aberrant cell shape induced in the yeast S. pombe by expression of transfected Drosophila cDNAs. These experiments show that elements that define cell shape are conserved throughout phylogeny and that a screen in yeast is a valuable tool for recovering heterologous cDNAs that encode cytoskeletal elements and the proteins that regulate them. In fly, we are identifying gene products that are necessary for myosin function by genetically recovering second site non-complementing loci and biochemically recovering proteins that bind to myosin. To date, our experiments identify ~30 loci that genetically interact with myosin and a kinase activity that phosphorylates myosin heavy chain and establish genetically, that the Rho signalling pathway is required in concert with nonmuscle myosin II for morphogenesis. We are also using manipulation studies to understand the forces that drive cellularization and morphogenesis. We show that both the amnioserosa and the leading edge of the lateral epidermis contribute to the movements of dorsal closure. Finally, we are examining the role these proteins play in movements that occur during wound healing.

Postdoctoral Fellow, Johns Hopkins University Medical School (Thomas D. Pollard, Advisor), 1982
Ph.D., University of Pennsylvania, 1979
B.A., University of Pennsylvania, 1973

Office Location: 4330 French Family Science Center, Science Drive, Duke University, Durham, NC 27
Office Phone: (919) 613-8157
Email Address: dkiehart@duke.edu
Web Page: http://www.biology.duke.edu/kiehartlab/
Additional Web Page: http://www.biology.duke.edu/kiehartlab/

Office Hours:

5:30 to 6:30 PM Mondays

Cell and Molecular Biology
Developmental Biology

Research Categories: Biophysical approaches to cellular, molecular and developmental biology

Current projects: Cytoskeleton and motor protein function in morphogenesis and wound healing, Light activated gene expression, Filopodia function in morphogenesis and wound healing, Protein complex function in hearing

Research Description: Our intellectual focus is on identifying determinants of cell shape that function during development and wound healing. We utilize novel biophysical strategies (in collaboration with Glenn Edwards' group in Physics and with Stephanos Venakide's and John Harer's groups in Mathematics) in concert with modern molecular genetic and reverse genetic approaches in Drosophila to explore the forces that are responsible for cell shape change and movements. We show that both the amnioserosa and a "supracellular purse string" in the leading edge of the lateral epidermis contribute to the movements of dorsal closure. Dorsal closure proceeds even if we ablate one (but NOT both!)of the tissues responsible for closure, indicating that this model cell (epithelial) sheet movement depends on redundant forces that in concert drive morphogenesis. We show that the magnitude of each force is significantly larger than their vector sum indicating that there is both potential for generating large forces and that successful morphogenesis requires that the forces applied be precisely balanced. We have also explored the molecules responsible for generating those movements. We showed that conventional nonmuscle myosin (myosin II) provides key contractile forces in different tissues where the supramolecular complexes that incorporate this motor protein are distinct. How molecular events are regulated such that large, opposing forces efficiently drive morphogenesis remains a mystery, but we are pursuing leads that point to two distinct pathways: the bidirectionally signaling integrin cell surface receptors and mechanically gated channels.

We are also pursuing the morphogenesis of actin-cytoskeleton based projections that are a key feature of a variety of cells, including those that are specialized for sensory reception in human vision and hearing. We have again turned to Drosophila as a model system where we study the morphogenesis of epidermal hairs and sensory bristles. Our work centers on an unconventional myosin (myosin VIIA) encoded by crinkled a gene that is required for the formation of epidermal hairs and bristles. We show that myosin VIIA is required for the coallescence of actin pre-hairs into the robust actin bundles that form the skeleton on which hairs and bristles can be built. In collaboration with Dan Eberl's lab (University of Iowa) we showed that myosin VIIA is also essential for fly hearing -- remarkably, its human homolog is also required for human hearing, even though the mechanisms of auditory sensory reception in these phylogenetically diverged systems are very different. We have begun to characterize myosin VIIA structurally using NMR of purified protein domains. With Jim Seller's lab at the NIH we have used fast time course kinetics and single molecule assays to analyze molecular function and show that this myosin VIIA is a processive motor. We are beginning to characterize the proteins that collaborate with both myosin II and myosin VIIA using biochemical strategies in vitro, yeast two hybrid approaches in vivo and genetic interaction strategies in fly.

Together, our experiments promise to reveal the nature of cytoskeletal function in cell shape determination for cell division and morphogenesis throughout development and organismal homeostasis.

Areas of Interest:
wound healing
motor protein structure and function
molecular structure
phylogeny of gene families

Recent Publications   (More Publications)   (search)

  1. Lu, H; Sokolow, A; Kiehart, DP; Edwards, GS, Quantifying dorsal closure in three dimensions., Molecular Biology of the Cell, vol. 27 no. 25 (December, 2016), pp. 3948-3955  [abs].
  2. Marston, DJ; Higgins, CD; Peters, KA; Cupp, TD; Dickinson, DJ; Pani, AM; Moore, RP; Cox, AH; Kiehart, DP; Goldstein, B, MRCK-1 Drives Apical Constriction in C.¬†elegans by Linking Developmental Patterning to Force Generation., Current Biology, vol. 26 no. 16 (August, 2016), pp. 2079-2089 [doi]  [abs].
  3. Lu, H; Sokolow, A; Kiehart, DP; Edwards, GS, Remodeling Tissue Interfaces and the Thermodynamics of Zipping during Dorsal Closure in Drosophila., Biophysical Journal, vol. 109 no. 11 (December, 2015), pp. 2406-2417 [doi]  [abs].
  4. Goldstein, B; Kiehart, DP, Moving Inward: Establishing the Mammalian Inner Cell Mass., Developmental Cell, vol. 34 no. 4 (August, 2015), pp. 385-386 [doi]  [abs].
  5. Kiehart, DP, Epithelial morphogenesis: apoptotic forces drive cell shape changes., Developmental Cell, vol. 32 no. 5 (March, 2015), pp. 532-533 [doi]  [abs].

Curriculum Vitae