Arabidopsis dynamin-like 2 that binds specifically to phosphatidylinositol 4-phosphate assembles into a high-molecular weight complex in vivo and in vitro.
Arabadopsis dynamin-like (ADL) 2, a member of the high-molecular weight (M(r)) dynamin family found in Arabidopsis, has been shown to be targeted to the plastid. In the chloroplast, most of the ADL2 was present in the fraction containing the envelope membranes when analyzed by suborganellar fractionation. Sucrose gradient and gel filtration experiments showed that when associated with membranes, ADL2 existed as a high-M(r) complex, whereas the soluble form existed as a monomer. The recombinant ADL2 expressed in Escherichia coli was present as a high-M(r) form and showed higher GTPase activity at a low NaCl concentration, whereas ADL2 existed as a low-M(r) form with a low level of GTPase activity at a high NaCl concentration. Electron microscopy studies revealed that the purified recombinant ADL2 formed spiral-coiled structures or rings. In the presence of guanosine-5'-O-(3-thio)triphosphate, these structures were transformed into a long rod structure. In contrast, in the presence of GDP, these structures disassembled into oligomers that were shown to be tetramer with 4-fold symmetry. Finally, a lipid-binding assay revealed that recombinant ADL2 purified from E. coli bound specifically to phosphatidylinositol 4-phosphate. Together, these results demonstrated that the biochemical properties of ADL2 were very similar to those of dynamin and other related proteins. Based on this similarity, we propose that ADL2 may be involved in vesicle formation at the chloroplast envelope membrane.
Kim
Y W
YW
Department of Life Science and Center for Plant Intracellular Trafficking, Pohang University of Science and Technology, Pohang, 790-784, Korea.
Park
D S
DS
Park
S C
SC
Kim
S H
SH
Cheong
G W
GW
Hwang
I
I
D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000378:metabolism*; D002736:Chloroplasts / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000378:metabolism; D005786:Gene Expression Regulation; D066298:In Vitro Techniques; D008970:Molecular Weight; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins; D012965:Sodium Chloride / Q000494:pharmacology
C114648:ADL2 protein, Arabidopsis; D029681:Arabidopsis Proteins; D018129:Phosphatidylinositol Phosphates; D010940:Plant Proteins; D011994:Recombinant Proteins; C037178:phosphatidylinositol 4-phosphate; D012965:Sodium Chloride; D020558:GTP Phosphohydrolases