Multiple Nuclear Localization Signals Mediate Nuclear Localization of the GATA Transcription Factor AreA

Hunter, Cameron C.; Siebert, Kendra S.; Downes, Damien J.; Wong, Koon Ho; Kreutzberger, Sara D.; Fraser, James A.; Clarke, David F.; Hynes, Michael J.; Davis, Meryl A.; Todd, Richard B.

doi:10.1128/ec.00040-14

Cited by 34 publications

(26 citation statements)

References 97 publications

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“…The PIKSRKE (NLS1), RKRP (NLS2) and LHGVVRPLSL (NLS3) sequences of AreA are three putative NLS sequences (Fig. A) that are conserved in its orthologues from Fusarium verticillioides , Fusarium oxysporum , Magnaporthe oryzae , N. crassa and A. nidulans (Hunter et al ., ). To determine their functions in the localization of AreA to the nucleus, we generated the AREA ΔNLS1 ‐GFP, AREA ΔNLS2 ‐GFP and AREA ΔNLS3 ‐GFP constructs and transformed them into the Δ areA mutant RH12.…”

Section: Resultsmentioning

confidence: 97%

“…However, deletion of either NLS1 or NLS2 of AreA had no obvious effect on its localization and function in F. graminearum . In A. nidulans , mutations at these two NLS sites also had no effect on the subcellular localization of AreA (Hunter et al ., ). However, AreA has a non‐canonical arginine‐based bipartite NLS sequence that is important for its nuclear localization.…”

Section: Discussionmentioning

confidence: 97%

“…Therefore, NLS3 must be the NLS sequence that is responsible for the subcellular localization of AreA in F. graminearum . In A. nidulans , mutations in the four R residues in this NLS also significantly reduced the localization of AreA to the nucleus (Hunter et al ., ). In F. graminearum , no detectable GFP signals were observed in the AREA ΔNLS3 ‐GFP transformant, even under NS conditions.…”

Section: Discussionmentioning

confidence: 97%

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The AreA transcription factor mediates the regulation of deoxynivalenol (DON) synthesis by ammonium and cyclic adenosine monophosphate (cAMP) signalling in Fusarium graminearum

Hou

Jiang

Zheng

et al. 2015

Molecular Plant Pathology

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Deoxynivalenol (DON), a trichothecene mycotoxin produced by Fusarium graminearum, is harmful to humans and animals. Because different nitrogen sources are known to have opposite effects on DON production, in this study, we characterized the regulatory mechanisms of the AREA transcription factor in trichothecene biosynthesis. The ΔareA mutant showed significantly reduced vegetative growth and DON production in cultures inoculated with hyphae. Suppression of TRI gene expression and DON production by ammonium were diminished in the ΔareA mutant. The deletion of AREA also affected the stimulatory effects of arginine on DON biosynthesis. The AreA-green fluorescent protein (GFP) fusion complemented the ΔareA mutant, and its localization to the nucleus was enhanced under nitrogen starvation conditions. Site-directed mutagenesis showed that the conserved predicted protein kinase A (PKA) phosphorylation site S874 was important for AreA function, indicating that AreA may be a downstream target of the cyclic adenosine monophosphate (cAMP)-PKA pathway, which is known to regulate DON production. We also showed that AreA interacted with Tri10 in co-immunoprecipitation assays. The interaction of AreA with Tri10 is probably related to its role in the regulation of TRI gene expression. Interestingly, the ΔareA mutant showed significantly reduced PKA activity and expression of all three predicted ammonium permease (MEP) genes, in particular MEP1, under low ammonium conditions. Taken together, our results show that AREA is involved in the regulation of DON production by ammonium suppression and the cAMP-PKA pathway. The AreA transcription factor may interact with Tri10 and control the expression and up-regulation of MEP genes.

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Section: Resultsmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 97%

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The AreA transcription factor mediates the regulation of deoxynivalenol (DON) synthesis by ammonium and cyclic adenosine monophosphate (cAMP) signalling in Fusarium graminearum

Hou

Jiang

Zheng

et al. 2015

Molecular Plant Pathology

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“…Nuclear localization is essential for controlling the transcription of protein-coding genes (24,25). The majority of NLSs contain basic amino acid sequences of 4-30 residues, which are too short to form independent structures, and typically exist within nuclear proteins.…”

Section: Discussionmentioning

confidence: 99%

A nuclear localization signal is required for the nuclear translocation of Fign and its microtubule‑severing function

Cheng

et al. 2020

Mol Med Report

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It is commonly known that the specific function of a given ATPase associated with diverse cellular activities protein (i.e., a member of the AAA superfamily of proteins) depends primarily on its subcellular location. The microtubule-severing protein fidgetin (Fign) possesses a nuclear localization signal (NLS) that facilitates its translocation to the nucleus, where its assembly is finalized; here, Fign contributes to the regulation of microtubule configuration by cutting and trimming microtubule polymers. In the present study, Fign was found to be a nuclear protein, whose N-terminal sequence (SSLKRKAFYM; residues 314-323) acts as an NLS. Following substitution (KR to NN; 317-318) or deletion (NT; 314-323) mutations within the NLS, Fign, which is predominantly expressed in the nucleus, was found to reside in the cytoplasm of transfected cells. Furthermore, Fign was found to have an essential role in microtubule severing by preferentially targeting highly-tyrosinated microtubules (tyr-MTs). Mutation of the Fign NLS did not affect its microtubule-severing function or the cleavage of tyr-MTs, but did affect the cellular distribution of the Fign protein itself. Taken altogether, an NLS for Fign was identified, and it was demonstrated that the basic amino acids K317 and R318 are necessary for regulating its entry into the nucleus, whereas an increase in Fign in the cytosol due to mutations of the NLS did not affect its cleavage function.

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“…Potentially, however, a protein can contain more than one signal sequence; indeed multiple NLSs have been described in many nuclear proteins, including GATA transcription factors 49 . Thus, when analyzing different fragments of the tested protein for Pd targeting, it is possible that more than one such fragment will be identified, indicating the presence of multiple PLSs which may act independently or synergistically.…”

Section: Discussionmentioning

confidence: 99%

Identification of Plasmodesmal Localization Sequences in Proteins <em>In Planta</em>

Cheng

Lazarowitz

Citovsky

2017

JoVE

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Plasmodesmata (Pd) are cell-to-cell connections that function as gateways through which small and large molecules are transported between plant cells. Whereas Pd transport of small molecules, such as ions and water, is presumed to occur passively, cell-to-cell transport of biological macromolecules, such proteins, most likely occurs via an active mechanism that involves specific targeting signals on the transported molecule. The scarcity of identified plasmodesmata (Pd) localization signals (PLSs) has severely restricted the understanding of protein-sorting pathways involved in plant cell-to-cell macromolecular transport and communication. From a wealth of plant endogenous and viral proteins known to traffic through Pd, only three PLSs have been reported to date, all of them from endogenous plant proteins. Thus, it is important to develop a reliable and systematic experimental strategy to identify a functional PLS sequence, that is both necessary and sufficient for Pd targeting, directly in the living plant cells. Here, we describe one such strategy using as a paradigm the cell-to-cell movement protein (MP) of the Tobacco mosaic virus (TMV). These experiments, that identified and characterized the first plant viral PLS, can be adapted for discovery of PLS sequences in most Pd-targeted proteins.

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Multiple Nuclear Localization Signals Mediate Nuclear Localization of the GATA Transcription Factor AreA

Cited by 34 publications

References 97 publications

The AreA transcription factor mediates the regulation of deoxynivalenol (DON) synthesis by ammonium and cyclic adenosine monophosphate (cAMP) signalling in Fusarium graminearum

The AreA transcription factor mediates the regulation of deoxynivalenol (DON) synthesis by ammonium and cyclic adenosine monophosphate (cAMP) signalling in Fusarium graminearum

A nuclear localization signal is required for the nuclear translocation of Fign and its microtubule‑severing function

Identification of Plasmodesmal Localization Sequences in Proteins <em>In Planta</em>

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