Continued studies will focus primarily on tenascin - the giant hexabrachion molecule in the extracellular matrix of tumors, healing wounds and specific embryonic tissues. We are already making transgenic mice that overexpress tenascin. We propose to make others that will be defective in hexabrachion assembly (monobrachion mice), and mice with depletion in tenascin secretion. We will determine the effects of overexpression, underexpression or defective hexabrachions on embryonic development, tumors and wound healing. A second project is a search for the tenascin gene in Drosophila and C. elegans. These species have highly developed genetic maps and large numbers of mutants, offering a powerful new approach to understanding the functions of tenascin. We will continue our studies of the cell biology and biochemistry of tenascin, to identify and characterize cell surface receptors for different domains of tenascin. A key tool for this project is the library of bacterial expression proteins that we have recently developed, providing large quantities of defined segments of the hexabrachion arm. We will use a similar approach to identify ECM molecules that bind to tenascin. Another continuing project is to characterize a new form of laminin that we identified during the purification of tenascin from cell culture supernatant. This new protein appears to have a variant A chain, a unique tissue distribution, and a cell adhesion activity quite different from any known laminin. Finally, we propose to expand our collaborative projects on the x-ray crystallography of tenascin. Our approach is to crystallize one domain at a time, using our PCR approach to make precisely defined bacterial expression proteins. The two FN-III domains that we have tried so far, the RGD domains from tenascin and fibronectin, have given excellent crystals, and high resolution x-ray diffraction studies are in progress. Additional domains are now proposed for future studies. An atomic resolution structure of the entire hexabrachion is a feasible goal.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37CA047056-20
Application #
2837626
Study Section
Special Emphasis Panel (NSS)
Program Officer
Mohla, Suresh
Project Start
1979-05-01
Project End
2000-11-30
Budget Start
1998-12-01
Budget End
1999-11-30
Support Year
20
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Duke University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
071723621
City
Durham
State
NC
Country
United States
Zip Code
27705
Shah, Riddhi; Ohashi, Tomoo; Erickson, Harold P et al. (2017) Spontaneous Unfolding-Refolding of Fibronectin Type III Domains Assayed by Thiol Exchange: THERMODYNAMIC STABILITY CORRELATES WITH RATES OF UNFOLDING RATHER THAN FOLDING. J Biol Chem 292:955-966
Erickson, Harold P (2017) Protein unfolding under isometric tension-what force can integrins generate, and can it unfold FNIII domains? Curr Opin Struct Biol 42:98-105
Albrecht, Elke; Norheim, Frode; Thiede, Bernd et al. (2015) Irisin - a myth rather than an exercise-inducible myokine. Sci Rep 5:8889
Giacomodonato, Mónica N; Noto Llana, Mariángeles; Aya Castañeda, María Del Rosario et al. (2014) AvrA effector protein of Salmonella enterica serovar Enteritidis is expressed and translocated in mesenteric lymph nodes at late stages of infection in mice. Microbiology 160:1191-9
Erickson, Harold P (2013) Irisin and FNDC5 in retrospect: An exercise hormone or a transmembrane receptor? Adipocyte 2:289-93
Fouda, Genevieve G; Jaeger, Frederick H; Amos, Joshua D et al. (2013) Tenascin-C is an innate broad-spectrum, HIV-1-neutralizing protein in breast milk. Proc Natl Acad Sci U S A 110:18220-5
Schumacher, Maria A; Chinnam, Nagababu; Ohashi, Tomoo et al. (2013) The structure of irisin reveals a novel intersubunit ?-sheet fibronectin type III (FNIII) dimer: implications for receptor activation. J Biol Chem 288:33738-44
Lemmon, Christopher A; Ohashi, Tomoo; Erickson, Harold P (2011) Probing the folded state of fibronectin type III domains in stretched fibrils by measuring buried cysteine accessibility. J Biol Chem 286:26375-82
Ohashi, Tomoo; Erickson, Harold P (2011) Fibronectin aggregation and assembly: the unfolding of the second fibronectin type III domain. J Biol Chem 286:39188-99
Ohashi, Tomoo; Erickson, Harold P (2009) Revisiting the mystery of fibronectin multimers: the fibronectin matrix is composed of fibronectin dimers cross-linked by non-covalent bonds. Matrix Biol 28:170-5

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