NDLI: Identification in vitro of a post-translational regulatory site in the hinge 1 region of Arabidopsis nitrate reductase.

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Identification in vitro of a post-translational regulatory site in the hinge 1 region of Arabidopsis nitrate reductase.

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Identification in vitro of a post-translational regulatory site in the hinge 1 region of Arabidopsis nitrate reductase.

Content Provider	PubMed Central
Author	Su, W. Huber, S. C. Crawford, N. M.
Abstract	Nitrate reductase (NR) is rapidly inactivated by phosphorylation of serine residues in response to loss of light or reduction in CO2 levels. To identify sites within NR protein that play a role in this post-translational regulation, a heterologous expression system and an in vitro inactivation assay for Arabidopsis NR were developed. Protein extracts containing NR kinases and inhibitor proteins were prepared from an NR-defective mutant that had lesions in both the NIA1 and NIA2 NR genes of Arabidopsis. Active NR protein was produced in a Pichia pastoris expression system. Incubation of these two preparations resulted in a Mg-ATP-dependent inactivation of NR that was reversed with EDTA. Mutant forms of NR were constructed, produced in P. pastoris, and tested in the in vitro inactivation assay. Six conserved serine residues in the hinge 1 region of NR, which separates the molybdenum cofactor and heme domains, were specifically targeted for mutagenesis because they are located in a potential regulatory region identified as a target for NR kinases in spinach. A change in Ser-534 to aspartate was found to block NR inactivation; changes in the other five serines had no effect. The aspartate that replaced Ser-534 did not appear to mimic a phosphorylated serine but simply prevented the NR from being inactivated. These results identify Ser-534, located in the hinge 1 of NR and conserved among higher plants NRs, as an essential site for post-translational regulation in vitro.
Starting Page	519
File Format	PDF
ISSN	1532298X
e-ISSN	1532298X
Journal	The Plant Cell
Issue Number	3
Volume Number	8
Language	English
Publisher Date	1996-03-01
Access Restriction	Open
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Cell Biology Plant Science

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Identification in vitro of a post-translational regulatory site in the hinge 1 region of Arabidopsis nitrate reductase.

Identification of Ser-543 as the major regulatory phosphorylation site in spinach leaf nitrate reductase.

Identification of the Arabidopsis CHL3 gene as the nitrate reductase structural gene NIA2.

Mechanism and importance of post‐translational regulation of nitrate reductase

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Identification of the 7-Hydroxymethyl Chlorophyll a Reductase of the Chlorophyll Cycle in Arabidopsis [W]

Ser-534 in the Hinge 1 Region of ArabidopsisNitrate Reductase Is Conditionally Required for Binding of 14-3-3 Proteins and in Vitro Inhibition

Identification of the Arabidopsis CHL3 gene as the nitrate reductase structural gene NIA2.

Differential Expression of the Two Arabidopsis Nitrate Reductase Genes 1

Identification in vitro of a post-translational regulatory site in the hinge 1 region of Arabidopsis nitrate reductase.

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