The isolation and characterization of mutant cells unable to perform the various steps of endocytosis may be the only way to fully understand this complex series of events and the interrelationship of endocytosis of intracellular membrane trafficking. Mutants in Chinese hamster ovary cells which have been isolated seem to fall into two general types, either receptor minus or those defective in endosomal acidification. The pleotropic phenotype of the acidification mutation has already provided new insights into the common mechanism involved in membrane trafficking. Recently, we have isolated a subline of the human derived hepatoblastoma cell line, HepG2, resistant to an asialoglycoprotein conjugate of amethopterin. Both parental and mutant cell lines were equally sensitive to amethopterin alone, indicating that drug resistance was not responsible for the survival of the mutant. Analysis of the status of the asialoglycoprotein receptor content and function in these cells indicated that there was a 60% loss of receptor activity with no concomitant loss of receptor protein expression. It is reasonable to assume that mutations other than of these two general classes would be lethal to the cell, and, therefore, we propose to isolate conditional mutants for various steps in the pathway. To target the selection for such mutants, it is proposed to use two cytotoxic conjugates as selective agents. By using two unrelated receptors that undergo endocytosis, it should be possible to avoid the selection of mutants that are receptor-specific. Once isolated, the defect in the intracellular pathway will be defined at both a functional and genetic level. The long-term objective of this proposal is to provide insight into the biochemical elements which control the endocytotic pathway.
| Dulyaninova, Natalya G; Ruiz, Penelope D; Gamble, Matthew J et al. (2018) S100A4 regulates macrophage invasion by distinct myosin-dependent and myosin-independent mechanisms. Mol Biol Cell 29:632-642 |
| Kakabadze, Zurab; Kakabadze, Ann; Chakhunashvili, David et al. (2018) Decellularized human placenta supports hepatic tissue and allows rescue in acute liver failure. Hepatology 67:1956-1969 |
| Rao, Lu; Hülsemann, Maren; Gennerich, Arne (2018) Combining Structure-Function and Single-Molecule Studies on Cytoplasmic Dynein. Methods Mol Biol 1665:53-89 |
| Gong, Zhenwei; Tasset, Inmaculada; Diaz, Antonio et al. (2018) Humanin is an endogenous activator of chaperone-mediated autophagy. J Cell Biol 217:635-647 |
| Kale, Abhijit; Ji, Zhejun; Kiparaki, Marianthi et al. (2018) Ribosomal Protein S12e Has a Distinct Function in Cell Competition. Dev Cell 44:42-55.e4 |
| Caballero, Benjamin; Wang, Yipeng; Diaz, Antonio et al. (2018) Interplay of pathogenic forms of human tau with different autophagic pathways. Aging Cell 17: |
| Akiyama, Matthew J; Agyemang, Linda; Arnsten, Julia H et al. (2018) Rationale, design, and methodology of a trial evaluating three models of care for HCV treatment among injection drug users on opioid agonist therapy. BMC Infect Dis 18:74 |
| Willis, Ian M (2018) Maf1 phenotypes and cell physiology. Biochim Biophys Acta Gene Regul Mech 1861:330-337 |
| Wang, Tony Y; Portincasa, Piero; Liu, Min et al. (2018) Mouse models of gallstone disease. Curr Opin Gastroenterol 34:59-70 |
| Hodge, Dayle Q; Cui, Jihong; Gamble, Matthew J et al. (2018) Histone Variant MacroH2A1 Plays an Isoform-Specific Role in Suppressing Epithelial-Mesenchymal Transition. Sci Rep 8:841 |
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