title: Interaction of D-lactate dehydrogenase protein 2 (Dld2p) with F-actin: implication for an alternative function of Dld2p.
abstract: D-Lactate dehydrogenase protein 2 [Yeast 15 (1999) 1377; Biochem. Biophys. Res. Commun. 295 (2002) 910] was initially identified as the actin interacting protein 2 (Aip2p) using a two-hybrid screen to search for proteins that interact with actin [Nat. Struct. Biol. 2 (1995) 28], but no other evidence indicating an interaction between Aip2p and actin cytoskeleton has been reported so far. During our search for the protein conformation modifying activity, we serendipitously identified Aip2p isolated from Saccharomyces cerevisiae as exhibiting an interaction with F-actin both in vitro and in vivo. Incubation with Aip2p facilitated the formation of the circular form of F-actin in vitro, which exhibited an aberrant trypsin susceptibility. Overexpression of Aip2p induced multi-buds in yeast cells, whereas reduced expression interfered with the formation of the cleavage furrow for the cell division, which was rescued by the introduction of wild-type Aip2p. While Aip2p-treated F-actin in the circular form was negligibly stained by rhodamine-labeled phalloidin (rhodamine-phalloidin) in vitro, rhodamine-phalloidin staining profiles in actin interacting protein 2 gene (AIP2)-modified cells suggested a correlation between the conformation of F-actin and the expression of Aip2p in vivo. AIP2-deleted cells became sensitive to osmotic conditions, a hallmark of actin dysfunction. Finally, immunoprecipitation of yeast cells using anti-Aip2p antibody demonstrated that Aip2p associates with actin. These properties suggest that Aip2p may interact with F-actin in vivo and play an important role in the yeast cell morphology.
authors.affiliation: Department of Cortical Function Disorders, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo, Japan.
authors.forename: Naomi S
authors.initials: NS
authors.lastname: Hachiya
authors.forename: Yuji
authors.initials: Y
authors.lastname: Sakasegawa
authors.forename: Akiko
authors.initials: A
authors.lastname: Jozuka
authors.forename: Shoichiro
authors.initials: S
authors.lastname: Tsukita
authors.forename: Kiyotoshi
authors.initials: K
authors.lastname: Kaneko
mesh_terms: D000199:Actins / Q000737:chemistry*; D002455:Cell Division; D003599:Cytoskeleton / Q000378:metabolism; D006639:Histidine / Q000737:chemistry; D007770:L-Lactate Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D042844:L-Lactate Dehydrogenase (Cytochrome); D045022:Lactate Dehydrogenases / Q000737:chemistry* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009995:Osmosis; D010590:Phalloidine / Q000737:chemistry; D011233:Precipitin Tests; D011485:Protein Binding; D011487:Protein Conformation; D012235:Rhodamines / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism; D014357:Trypsin / Q000737:chemistry; D020798:Two-Hybrid System Techniques
chemical_list: D000199:Actins; D012235:Rhodamines; D029701:Saccharomyces cerevisiae Proteins; D010590:Phalloidine; D006639:Histidine; D045022:Lactate Dehydrogenases; D007770:L-Lactate Dehydrogenase; C071195:D-lactate dehydrogenase; D042844:L-Lactate Dehydrogenase (Cytochrome); C487563:DLD2 protein, S cerevisiae; D014357:Trypsin