Interaction of D-lactate dehydrogenase protein 2 (Dld2p) with F-actin: Implication for an alternative function of Dld2p

Naomi S. Hachiya, Yuji Sakasegawa, Akiko Jozuka, Shoichiro Tsukita, Kiyotoshi Kaneko

Research output: Contribution to journalArticle

7 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)78-82
Number of pages5
JournalBiochemical and biophysical research communications
Volume319
Issue number1
DOIs
Publication statusPublished - 2004 Jun 18

Keywords

  • Actin interacting protein 2
  • Actin interacting protein 2 gene
  • D-Lactate dehydrogenase protein 2
  • F-actin
  • Rhodamine-phalloidin staining
  • Trypsin susceptibility assay
  • Yeast cell morphology

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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