Abstract
Small GTPase Rab36 is homologous to Rab34 with 56% amino acid sequence identity. Rab34 was characterized as a Golgi-associated Rab protein and regulates lysosomal positioning through interaction with RILP; however, the properties and functions of Rab36 have not been investigated. To investigate Rab36, we constructed EGFP-Rab36 wild type, the active GTP-bound mutant EGFP-Rab36Q116L and negative GDP-bound mutant EGFP-Rab36T71N. Expression of EGFP-Rab36 wild type revealed that Rab36 co-localized with Golgi markers GM130, Syntaxin 5 and TGN46 in Hela cells, indicating Rab36 is associated with Golgi apparatus. Over-expression of Rab36 induced late endosome and lysosome clustering around the Golgi apparatus, marked by LBPA, CD63, Lamp1 and Lamp2, without effects on early endosomal compartment marked by EEA1. GST-pulldown assay demonstrated that Rab36 can also interact with RILP. In addition, the binding region for Rab36 is in the C-terminal region (aa199-401) of RILP. Our data suggested that Rab36 may regulate the spatial distribution of late endosomes and lysosomes through a similar mechanism to Rab34.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing / genetics
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Adaptor Proteins, Signal Transducing / metabolism
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Amino Acid Substitution
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Antigens, CD / genetics
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Antigens, CD / metabolism
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Autoantigens / genetics
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Autoantigens / metabolism
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Endosomes / genetics
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Endosomes / metabolism*
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Golgi Apparatus / genetics
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Golgi Apparatus / metabolism*
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HeLa Cells
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Humans
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Lysosomal Membrane Proteins / genetics
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Lysosomal Membrane Proteins / metabolism
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Lysosomal-Associated Membrane Protein 2
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Lysosomes / genetics
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Lysosomes / metabolism*
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Membrane Glycoproteins / genetics
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Membrane Glycoproteins / metabolism
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Membrane Proteins / genetics
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Membrane Proteins / metabolism
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Mutation, Missense
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Platelet Membrane Glycoproteins / genetics
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Platelet Membrane Glycoproteins / metabolism
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Qa-SNARE Proteins / genetics
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Qa-SNARE Proteins / metabolism
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Tetraspanin 30
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rab GTP-Binding Proteins / genetics
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rab GTP-Binding Proteins / metabolism*
Substances
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Adaptor Proteins, Signal Transducing
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Antigens, CD
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Autoantigens
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CD63 protein, human
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Golgin subfamily A member 2
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LAMP1 protein, human
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LAMP2 protein, human
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Lysosomal-Associated Membrane Protein 2
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Lysosomal Membrane Proteins
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Membrane Glycoproteins
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Membrane Proteins
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Platelet Membrane Glycoproteins
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Qa-SNARE Proteins
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RILP protein, human
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TGOLN2 protein, human
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Tetraspanin 30
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RAB39A protein, human
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Rab36 protein, human
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rab GTP-Binding Proteins