NBP35 interacts with DRE2 in the maturation of cytosolic iron-sulfur proteins in Arabidopsis thaliana

Bastow, Emma L., Bych, Katrine, Crack, Jason C., Le Brun, Nick E. and Balk, Janneke (2017) NBP35 interacts with DRE2 in the maturation of cytosolic iron-sulfur proteins in Arabidopsis thaliana. The Plant Journal, 89 (3). 590–600. ISSN 0960-7412

[img] PDF (Accepted manuscript) - Submitted Version
Restricted to Repository staff only until 31 December 2099.

Download (488kB) | Request a copy
    [img]
    Preview
    PDF (Published manuscript) - Published Version
    Available under License Creative Commons Attribution.

    Download (854kB) | Preview

      Abstract

      Proteins of the cytosolic pathway for iron-sulfur (FeS) cluster assembly are conserved, except that plants lack a gene for CFD1 (Cytosolic FeS cluster Deficient 1). This poses the question of how NBP35 (Nucleotide-Binding Protein 35 kDa), the heteromeric partner of CFD1 in metazoa, functions on its own in plants. Firstly, we created viable mutant alleles of NBP35 in Arabidopsis to overcome embryo lethality of previously reported knockout mutations. RNAi knockdown lines with less than 30% NBP35 protein surprisingly showed no developmental or biochemical differences to wild type. Substitution of Cys14 to Ala, which destabilized the N-terminal Fe4S4 cluster in vitro, caused mild growth defects and a significant decrease in the activity of cytosolic FeS enzymes such as aconitase and aldehyde oxidases. The DNA glycosylase ROS1 was only partially decreased in activity and xanthine dehydrogenase not at all. Plants with strongly depleted NBP35 protein in combination with Cys14 to Ala substitution had distorted leaf development and decreased FeS enzyme activities. To find protein interaction partners of NBP35, a yeast two-hybrid screen was carried out which identified NBP35 and DRE2 (Derepressed for Ribosomal protein S14 Expression). NBP35 is known to form a dimer, and DRE2 acts upstream in the cytosolic FeS protein assembly pathway. The NBP35-DRE2 interaction was not disrupted by Cys14 to Ala substitution. Our results show that NBP35 has a function in the maturation of FeS proteins that is conserved in plants, and is closely allied to the function of DRE2.

      Item Type: Article
      Uncontrolled Keywords: fe–s cofactor,cytosol,leaf development,aconitase,aldehyde oxidase,dna methylation,ciapin1,yeast-two-hybrid,arabidopsis thaliana
      Faculty \ School: Faculty of Science > School of Chemistry
      Faculty of Science > School of Biological Sciences
      Related URLs:
      Depositing User: Pure Connector
      Date Deposited: 08 Nov 2016 15:00
      Last Modified: 12 Jun 2019 01:08
      URI: https://ueaeprints.uea.ac.uk/id/eprint/61265
      DOI: 10.1111/tpj.13409

      Actions (login required)

      View Item