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DNA-protein affinity precipitation to identify proteins interacting with the atpT promoter in Synechocystis sp. PCC 6803

ABSTRACT: The small protein AtpΘ (encoded by gene atpT) identified in cyanobacteria becomes maximum expressed during low-energy conditions such as during darkness. In the model cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis 6803), AtpΘ was demonstrated to inhibit the ATP hydrolysis activity of the F0F1 ATP synthase. Here, the regulation of atpT gene expression was studied in greater detail. The darkness-induced activation of the atpT promoter indicated the existence of regulatory factors. To identify such factors, DNA-protein affinity precipitation was performed using biotinylated DNA fragments. These fragments contained either the promoter-5’UTR (PatpT66-5’UTR) as bait or, as negative control, only the 5’UTR (atpT 5’UTR). Each DNA fragment was incubated with total protein samples isolated from exponential phase Synechocystis 6803 which had been kept in the light or 12 h in darkness prior to protein preparation. The resulting protein samples were then analyzed by mass spectrometry.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Synechocystis Sp. Pcc 6803

SUBMITTER: Sandra Maass

LAB HEAD: Dörte Becher

PROVIDER: PXD027801 | Pride | 2022-04-14

REPOSITORIES: Pride

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