The isolation of Saccharomyces cerevisiae nuclear membranes with nuclease and high-salt treatment

Mann, Karlheinz; Mecke, Dieter

doi:10.1016/0005-2736(82)90169-9

Cited by 8 publications

(2 citation statements)

References 27 publications

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“…Nuclear membranes were prepared from the wild-type S. cereisiae cells as described in [28]. For preparation of liposomes from nuclear membranes, the membranes were extracted with 100 % chloroform and the liposomes were prepared.…”

Section: Preparation Of Nuclear Membranesmentioning

confidence: 99%

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Inhibitory effects of acidic phospholipids on the binding of origin-recognition complex to origin DNA

Lee

Makise

Takenaka

et al. 2002

Biochemical Journal

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Origin-recognition complex (ORC), a candidate initiator of chromosomal DNA replication in eukaryotes, shares certain biochemical characteristics with DnaA, the initiator of chromosomal DNA replication in prokaryotes. These similarities include origin-specific DNA binding, ATP binding and ATPase activity. DnaA interacts with acidic phospholipids, such as cardiolipin, and its activity is regulated by these phospholipids. In this study, we examined whether Saccharomyces cerevisiae ORC also interacts with phospholipids. Among the various phospholipids tested, ORC was found to bind specifically to cardiolipin. This binding was inhibited by excess concentrations of salts but unaffected by ATP, adenosine 5′-[γ-thio]triphosphate or the origin DNA. Cardiolipin weakly inhibited the ATP-binding activity of ORC, whereas it strongly inhibited ORC binding to origin DNA. Acidic phospholipids other than cardiolipin (phosphatidylglycerol and phosphatidylinositol) weakly inhibited ORC binding to origin DNA. Furthermore, total phospholipids extracted from yeast nuclear membranes inhibited ORC binding to origin DNA. We consider that phospholipids may modulate initiation of DNA replication in eukaryotes in a similar manner to that found in prokaryotes.

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Section: Preparation Of Nuclear Membranesmentioning

confidence: 99%

“…in nuclear membranes. According to a previous study [28], nuclear-membrane fractions were prepared from wild-type yeast cells and the amount of ORC in the fraction determined by immunoblotting. Approx.…”

Section: Effect Of CL On the Interaction Between Orc And Origin Dnamentioning

confidence: 99%