Yeast importin- is required for nuclear import of the Mig2 repressorReport as inadecuate




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BMC Cell Biology

, 13:31

Nuclear function and transport

Abstract

BackgroundMig2 has been described as a transcriptional factor that in the absence of Mig1 protein is required for glucose repression of the SUC2 gene. Recently it has been reported that Mig2 has two different subcellular localizations. In high-glucose conditions it is a nuclear modulator of several Mig1-regulated genes, but in low-glucose most of the Mig2 protein accumulates in mitochondria. Thus, the Mig2 protein enters and leaves the nucleus in a glucose regulated manner. However, the mechanism by which Mig2 enters into the nucleus was unknown until now.

ResultsHere, we report that the Mig2 protein is an import substrate of the carrier Kap95 importin-. The Mig2 nuclear import mechanism bypasses the requirement for Kap60 importin- as an adaptor protein, since Mig2 directly binds to Kap95 in the presence of Gsp1GDP. We also show that the Mig2 nuclear import and the binding of Mig2 with Kap95 are not glucose-dependent processes and require a basic NLS motif, located between lysine-32 and arginine-37. Mig2 interaction with Kap95 was assessed in vitro using purified proteins, demonstrating that importin-, together with the GTP-binding protein Gsp1, is able to mediate efficient Mig2-Kap95 interaction in the absence of the importin- Kap60. It was also demonstrated, that the directionality of Mig2 transport is regulated by association with the small GTPase Gsp1 in the GDP- or GTP-bound forms, which promote cargo recognition and release, respectively.

ConclusionsThe Mig2 protein accumulates in the nucleus through a Kap95 and NLS-dependent nuclear import pathway, which is independent of importin- in Saccharomyces cerevisiae.

AbbreviationsNLSNuclear localization sequence

GSTGlutathione S-transferase

GFPGreen fluorescent protein

RFPRed fluorescent protein

DAPI4,6-Diamino-2-phenylindole.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2121-13-31 contains supplementary material, which is available to authorized users.

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Author: AlejandraFernndez-Cid - MontserratVega - PilarHerrero - FernandoMoreno

Source: https://link.springer.com/



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