Split-ubiquitin yeast two-hybrid interaction reveals a novel interaction between a natural resistance associated macrophage protein and a membrane bound thioredoxin ... View Full Text


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Article Info

DATE

2016-08-17

AUTHORS

Ananya Marik, Haraprasad Naiya, Madhumanti Das, Gairik Mukherjee, Soumalee Basu, Chinmay Saha, Rajdeep Chowdhury, Kankan Bhattacharyya, Anindita Seal

ABSTRACT

Natural resistance associated macrophage proteins (NRAMPs) are evolutionarily conserved metal transporters involved in the transport of essential and nonessential metals in plants. Fifty protein interactors of a Brassica juncea NRAMP protein was identified by a Split-Ubiquitin Yeast-Two-Hybrid screen. The interactors were predicted to function as components of stress response, signaling, development, RNA binding and processing. BjNRAMP4.1 interactors were particularly enriched in proteins taking part in photosynthetic or light regulated processes, or proteins predicted to be localized in plastid/chloroplast. Further, many interactors also had a suggested role in cellular redox regulation. Among these, the interaction of a photosynthesis-related thioredoxin, homologous to Arabidopsis HCF164 (High-chlorophyll fluorescence164) was studied in detail. Homology modeling of BjNRAMP4.1 suggested that it could be redox regulated by BjHCF164. In yeast, the interaction between the two proteins was found to increase in response to metal deficiency; Mn excess and exogenous thiol. Excess Mn also increased the interaction in planta and led to greater accumulation of the complex at the root apoplast. Network analysis of Arabidopsis homologs of BjNRAMP4.1 interactors showed enrichment of many protein components, central to chloroplastic/cellular ROS signaling. BjNRAMP4.1 interacted with BjHCF164 at the root membrane and also in the chloroplast in accordance with its proposed function related to photosynthesis, indicating that this interaction occurred at different sub-cellular locations depending on the tissue. This may serve as a link between metal homeostasis and chloroplastic/cellular ROS through protein–protein interaction. More... »

PAGES

519-537

References to SciGraph publications

  • 2014-10-19. Crystal structure of a SLC11 (NRAMP) transporter reveals the basis for transition-metal ion transport in NATURE STRUCTURAL & MOLECULAR BIOLOGY
  • 2013-09-19. A node-based switch for preferential distribution of manganese in rice in NATURE COMMUNICATIONS
  • 1997-07. Cloning and characterization of a mammalian proton-coupled metal-ion transporter in NATURE
  • 2011-09-23. Nutritional Immunity: Homology Modeling of Nramp Metal Import in CURRENT TOPICS IN INNATE IMMUNITY II
  • 2006-10-25. Mining the Arabidopsis thaliana genome for highly-divergent seven transmembrane receptors in GENOME BIOLOGY
  • 2012-02-24. Characterizing the role of rice NRAMP5 in Manganese, Iron and Cadmium Transport in SCIENTIFIC REPORTS
  • 2010-12-09. Interaction of the heterotrimeric G protein alpha subunit SSG-1 of Sporothrix schenckii with proteins related to stress response and fungal pathogenicity using a yeast two-hybrid assay in BMC MICROBIOLOGY
  • 1992-12. The expression of a rab-related gene, rab18, is induced by abscisic acid during the cold acclimation process of Arabidopsis thaliana (L.) Heynh in PLANT MOLECULAR BIOLOGY
  • 2008-01-22. Protocol: Streamline cloning of genes into binary vectors in Agrobacterium via the Gateway® TOPO vector system in PLANT METHODS
  • 2005-12. Uranium Accumulation of Crop Plants Enhanced by Citric Acid in ENVIRONMENTAL GEOCHEMISTRY AND HEALTH
  • 2011-03-11. Isolation and expression analysis of partial sequences of heavy metal transporters from Brassica juncea by coupling high throughput cloning with a molecular fingerprinting technique in PLANTA
  • Identifiers

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    PUBMED

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