title: Deciphering the role of Atg5 in nucleotide dependent interaction of Rab33B with the dimeric complex, Atg5-Atg16L1.
abstract: Autophagy is a lysosomal degradation pathway that degrades cytosolic constituents, including whole organelles and intracellular pathogens. Previous studies on various autophagy related genes revealed the importance of the Atg12-Atg5-Atg16 complex in autophagy. Atg16L1 is an effector of Golgi-resident Rab33B and the molecular mechanism of the interaction of Rab33B with either Atg16L1 or in complex with Atg5 is still elusive. In the current study, using the pull down and calorimetric approaches, we have dissected the molecular insights into the interaction of Rab33B with different regions of mouse Atg16L1 as well as with the dimeric complex, Atg5-mAtg16L1. Our in vitro observation suggests that Atg5 is pre-requisite for the augmented nucleotide dependent interaction of Rab33B with the dimeric complex, Atg5-Atg16L1. Moreover, the results reported here suggest that Arg-24 of Atg16L1 is crucial for its interaction with Atg5 which will have further implication in the binding of the dimeric complex to Rab33B.
authors.affiliation: Department of Biological Sciences, Indian Institute of Science Education and Research Bhopal-462023, India.
authors.forename: Mintu
authors.initials: M
authors.lastname: Chandra
authors.affiliation: University of Waterloo, Waterloo Institute for Nanotechnology, Waterloo, ON, Canada.
authors.forename: Runjhun
authors.initials: R
authors.lastname: Saran
authors.affiliation: Department of Biological Sciences, Indian Institute of Science Education and Research Bhopal-462023, India. Electronic address: sunando@iiserb.ac.in.
authors.forename: Sunando
authors.initials: S
authors.lastname: Datta
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001343:Autophagy; D000071187:Autophagy-Related Protein 5; D000071183:Autophagy-Related Proteins; D002151:Calorimetry; D002352:Carrier Proteins / Q000737:chemistry / Q000502:physiology*; D003001:Cloning, Molecular; D005982:Glutathione Transferase / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000737:chemistry; D006160:Guanosine Triphosphate / Q000737:chemistry; D006801:Humans; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000502:physiology*; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009711:Nucleotides / Q000737:chemistry*; D011485:Protein Binding; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013816:Thermodynamics; D020691:rab GTP-Binding Proteins / Q000737:chemistry / Q000502:physiology*
chemical_list: C532820:Atg16l1 protein, mouse; C495773:Atg5 protein, mouse; D000071187:Autophagy-Related Protein 5; D000071183:Autophagy-Related Proteins; D002352:Carrier Proteins; D008869:Microtubule-Associated Proteins; D009711:Nucleotides; D011994:Recombinant Proteins; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate; D005982:Glutathione Transferase; C486397:RAB33B protein, human; D020691:rab GTP-Binding Proteins
keywords: Autophagy; GST-pull down; Rab GTPase; Size exclusion chromatography; Thermodynamics