The National Institute of Diabetes & Digestive & Kidney Diseases

Our laboratory is interested in membrane and protein recycling within eukaryotic cells. The approach we are taking is to examine the structure and function of proteins involved in these processes by transmission electron microscopy, including cyro-electron microscopy, and by biochemical methods. Presently, we are examining a 100 kDa GTPase called dynamin, which is involved in the constriction and fission of clathrin-coated pits from plasma membrane during receptor-mediated endocytosis and during membrane retrieval in nerve terminals. We have shown that dynamin self-assembles into spirals and onto lipid bilayers forming dynamin decorated lipid tubes that undergo a large conformational change upon GTP addition resulting in membrane constriction. To further understand the mechanism of dynamin-induced membrane constriction, we have solved the 3-dimensional structure of dynamin in the non-constricted and constricted states by high-resolution electron microscopy and image processing methods. This work suggests a GTP-induced conformational change within the dynamin oligomer is responsible for constriction. Additional dynamin family members have been implicated in numerous fundamental cellular processes, including other membrane fission events, anti-viral activity, cell plate formation and chloroplast biogenesis. We are currently examining the effects of GTP on these proteins to determine if a common mechanism of action exists for all dynamin family members.

1. Nie Z, Hirsch DS, Luo R, Jian X, Stauffer S, Cremesti A, Andrade J, Lebowitz J, Marino M, Ahvazi B, Hinshaw JE, Randazzo PA A BAR domain in the N terminus of the Arf GAP ASAP1 affects membrane structure and trafficking of epidermal growth factor receptor. Curr Biol (16): 130-9, 2006. [Full Text/Abstract]

2. Naylor K, Ingerman E, Okreglak V, Marino M, Hinshaw JE, Nunnari J Mdv1 interacts with assembled dnm1 to promote mitochondrial division. J Biol Chem (281): 2177-83, 2006. [Full Text/Abstract]

3. Kochs G, Reichelt M, Danino D, Hinshaw JE, Haller O Assay and functional analysis of dynamin-like Mx proteins. Methods Enzymol (404): 632-43, 2005. [Full Text/Abstract]

4. Ingerman E, Perkins EM, Marino M, Mears JA, McCaffery JM, Hinshaw JE, Nunnari J Dnm1 forms spirals that are structurally tailored to fit mitochondria. J Cell Biol (170): 1021-7, 2005. [Full Text/Abstract]

5. Hickman AB, Perez ZN, Zhou L, Musingarimi P, Ghirlando R, Hinshaw JE, Craig NL, Dyda F Molecular architecture of a eukaryotic DNA transposase. Nat Struct Mol Biol (12): 715-21, 2005. [Full Text/Abstract]

6. Danino D, Moon KH, Hinshaw JE Rapid constriction of lipid bilayers by the mechanochemical enzyme dynamin. J Struct Biol (147): 259-67, 2004. [Full Text/Abstract]

7. Chen YJ Zhang P Egelman EH Hinshaw JE The stalk region of dynamin drives the constriction of dynamin tubes. Nat Struct Mol Biol (11): 574-5, 2004. [Full Text/Abstract]

8. Hinshaw JE Milligan RA Nuclear pore complexes exceeding eightfold rotational symmetry. J Struct Biol (141): 259-68, 2003. [Full Text/Abstract]

9. Danino D Hinshaw JE Dynamin family of mechanoenzymes. Curr Opin Cell Biol (13): 454-60, 2001. [Full Text/Abstract]

10. Zhang P Hinshaw JE Three-dimensional reconstruction of dynamin in the constricted state. Nat Cell Biol (3): 922-6, 2001. [Full Text/Abstract]

11. Hinshaw JE Dynamin and its role in membrane fission. Annu Rev Cell Dev Biol (16): 483-519, 2000. [Full Text/Abstract]

12. Hinshaw JE Dynamin spirals. Curr Opin Struct Biol (9): 260-7, 1999. [Full Text/Abstract]

13. Sweitzer SM Hinshaw JE Dynamin undergoes a GTP-dependent conformational change causing vesiculation. Cell (93): 1021-9, 1998. [Full Text/Abstract]

14. Carr JF Hinshaw JE Dynamin assembles into spirals under physiological salt conditions upon the addition of GDP and gamma-phosphate analogues. J Biol Chem (272): 28030-5, 1997. [Full Text/Abstract]

15. Warnock DE Hinshaw JE Schmid SL Dynamin self-assembly stimulates its GTPase activity. J Biol Chem (271): 22310-4, 1996. [Full Text/Abstract]

16. Baba T, Damke H, Hinshaw JE, Ikeda K, Schmid SL, Warnock DE Role of dynamin in clathrin-coated vesicle formation. Cold Spring Harb Symp Quant Biol (60): 235-42, 1995. [Full Text/Abstract]

17. Hinshaw JE Architecture of the nuclear pore complex and its involvement in nucleocytoplasmic transport. Biochem Pharmacol (47): 15-20, 1994. [Full Text/Abstract]

18. Hinshaw JE Carragher BO Milligan RA Architecture and design of the nuclear pore complex. Cell (69): 1133-41, 1992. [Full Text/Abstract]