DF/HCC Program Affiliation

Angiogenesis, Invasion, and Metastasis

Research Abstract

Our objective is to understand the molecular mechanisms governing the specificity and regulation of macro molecular traffic mediated by clathrin-coated pits and coated vesicles, a major apparatus that the cell uses to move receptors and ligands into and through the cell. These organelles undergo continual, rapid cycles of assembly and disassembly leading to the selective endocytosis of molecules such as growth factor receptors and neurotransmitters, immunoglobulins, low density lipoproteins and certain viruses; the traffic of the mannose 6-phosphate receptor; 7-transmembrane, g-coupled receptors and the secretion of insulin and ACTH.

Clathrin, the building block of the coat, organizes itself into basket-shaped lattices, providing structural stability for the vesicles, and, possibly, the driving force for internalization of segments of membranes. APs, the most prominent and ubiquitous of the proteins associated with the clathrin coat, are related heterotetrameric assemblies, whose detailed polypeptide composition depends on their intracellular location: AP-1 are complexes associated with coats at the trans-Golgi network and AP-2 are the complexes associated with coats at the plasma membrane. These APs interact with receptors, clathrin, and other membrane-associated proteins, and they appear to serve as critical regulatory junctions in coated vesicle activities.

We are currently examining the high resolution structure (by x-rays and electron microscopy) of clathrin and its partner proteins; how clathrin interacts with its AP complexes in order to form coats; the mechanism that allows the specific and regulated association of AP-1 and AP-2 complexes with the trans-Golgi network or the plasma membrane; and how an AP complex recognizes the membrane receptor that is specifically recruited into a coated pit. With these biochemical and cell biological studies we expect to obtain a framework for analyzing some of the molecular contacts and switches that participate in the regulation, availability and intracellular traffic of the many molecules involved in signal transduction, immune response, lipid homeostasis and cell-cell recognition.

Publications

• Fotin A, Cheng Y, Grigorieff N, Walz T, Harrison SC, Kirchhausen T.Structure of an auxilin-bound clathrin coat and its implications for the mechanism of uncoating.Nature 2004 Dec 2;432(7017):649-53.
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• Ehrlich M, Boll W, Van Oijen A, Hariharan R, Chandran K, Nibert ML, Kirchhausen T.Endocytosis by random initiation and stabilization of clathrin-coated pits.Cell 2004 Sep 3;118(5):591-605.
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• Kirchhausen T, Macia E, Pelish HE.Use of dynasore, the small molecule inhibitor of dynamin, in the regulation of endocytosis.Methods Enzymol 2008;438:77-93.
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• Boucrot E, Kirchhausen T.Mammalian Cells Change Volume during Mitosis.PLoS ONE 2008;3(1):e1477.
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• Nieland TJ, Shaw JT, Jaipuri FA, Duffner JL, Koehler AN, Banakos S, Zannis VI, Kirchhausen T, Krieger M.Identification of the molecular target of small molecule inhibitors of HDL receptor SR-BI activity.Biochemistry 2008 Jan 8;47(1):460-72.
  18067275
• Veiga E, Guttman JA, Bonazzi M, Boucrot E, Toledo-Arana A, Lin AE, Enninga J, Pizarro-Cerda J, Finlay BB, Kirchhausen T, Cossart P.Invasive and adherent bacterial pathogens co-Opt host clathrin for infection.Cell Host Microbe 2007 Nov 15;2(5):340-51.
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• Saffarian S, Kirchhausen T.Differential evanescence nanometry: live-cell fluorescence measurements with 10-nm axial resolution on the plasma membrane.Biophys J 2008 Mar 15;94(6):2333-42.
  17993495
• Rapoport I, Boll W, Yu A, Bocking T, Kirchhausen T.A motif in the clathrin heavy chain required for the hsc70/auxilin uncoating reaction.Mol Biol Cell 2008 Jan;19(1):405-13.
  17978091
• Li J, Peters PJ, Bai M, Dai J, Bos E, Kirchhausen T, Kandror KV, Hsu VW.An ACAP1-containing clathrin coat complex for endocytic recycling.J Cell Biol 2007 Jul 30;178(3):453-64.
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• Kirchhausen T.Making COPII coats.Cell 2007 Jun 29;129(7):1251-2.
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• Jiang S, Zhou X, Kirchhausen T, Wong ST.Tracking molecular particles in live cells using fuzzy rule-based system.Cytometry A 2007 Aug;71(8):576-84.
  17542029
• Wang J, Sun HQ, Macia E, Kirchhausen T, Watson H, Bonifacino JS, Yin HL.PI4P promotes the recruitment of the GGA adaptor proteins to the trans-Golgi network and regulates their recognition of the ubiquitin sorting signal.Mol Biol Cell 2007 Jul;18(7):2646-
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• Yu A, Rual JF, Tamai K, Harada Y, Vidal M, He X, Kirchhausen T.Association of Dishevelled with the clathrin AP-2 adaptor is required for Frizzled endocytosis and planar cell polarity signaling.Dev Cell 2007 Jan;12(1):129-41.
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• Benedito AB, Lehtinen M, Massol R, Lopes UG, Kirchhausen T, Rao A, Bonni A.The transcription factor NFAT3 mediates neuronal survival.J Biol Chem 2005 Jan 28;280(4):2818-25.
  15537643
• Rohatgi R, Ma L, Miki H, Lopez M, Kirchhausen T, Takenawa T, Kirschner MW.The interaction between N-WASP and the Arp2/3 complex links Cdc42-dependent signals to actin assembly.Cell 1999 Apr 16;97(2):221-31.
  10219243
• ter Haar E, Musacchio A, Harrison SC, Kirchhausen T.Atomic structure of clathrin: a beta propeller terminal domain joins an alpha zigzag linker.Cell 1998 Nov 13;95(4):563-73.
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• Tong X, Boll W, Kirchhausen T, Howley PM.Interaction of the bovine papillomavirus E6 protein with the clathrin adaptor complex AP-1.J Virol 1998 Jan;72(1):476-82.
  9420248