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Gene Review

NIT1  -  nitrilase 1

Arabidopsis thaliana

Synonyms: ATNIT1, NITI, NITRILE AMINOHYDROLASE
 
 
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Disease relevance of NIT1

  • Translational fusions placing the nit1 full-length promoter and a series of 5'-deletion fragments in front of the uidA gene encoding beta-glucuronidase (GUS) were used for Agrobacterium tumefaciens-mediated transformation of Nicotiana tabacum [1].
  • From an Arabidopsis thaliana cDNA expression library, a cDNA clone was isolated, characterized and sequenced which, at the amino acid level, resembled the Klebsiella ozaenae bromoxynil nitrilase encoded by the bxn gene [2].
  • The nitrilase AtNIT1 from Arabidopsis thaliana was overexpressed in Escherichia coli with an N-terminal His6 tag and purified by zinc chelate affinity chromatography in a single step almost to homogeneity in a 68% yield with a specific activity of 34.1 U.mg-1 [3].
 

High impact information on NIT1

 

Biological context of NIT1

 

Anatomical context of NIT1

  • To probe the capacity of ths enzyme under physiological conditions in vivo, the cDNA PM255, encoding nitrilase II, was stably integrated into the genome of Nicotiana tabacum by direct protoplast transformation under the control of the CaMV-35S promotor [9].
 

Associations of NIT1 with chemical compounds

  • NIT4 may play a role in cyanide detoxification during ethylene biosynthesis because extracts from senescent leaves of A. thaliana showed higher Ala(CN) hydratase/nitrilase activities than extracts from nonsenescent tissue [10].
  • Collectively, these data suggest that the elevated levels of IAM in seeds and germinating seedlings result from nitrilase action on indole-3-acetonitrile, a metabolite produced in the plants presumably from glucobrassicin turnover [11].
  • The mRNA levels of a diverse array of gene products increases during leaf senescence, including a protease, a ribosomal protein, two cinnamyl alcohol dehydrogenases, a nitrilase and glyoxalase II [12].
  • The stability of AtNIT1 could be significantly improved by the addition of dithiothreitol and EDTA [3].
 

Other interactions of NIT1

  • Genomic Southern hybridization, among other experiments, strongly suggests that tobacco has NIT4 homologs but not NIT1 to NIT3 homologs [7].
  • Three (NIT1, NIT2 and NIT3) of the four genes have high similarity [7].
  • The Arabidopsis thaliana isogene NIT4 and its orthologs in tobacco encode beta-cyano-L-alanine hydratase/nitrilase [10].
  • Transcript levels known to be involved in IAA/glucosinolate synthesis and metabolism (nitrilase and myrosinase) were examined and both showed developmental regulation, while only nitrilase mRNA levels differed between wild type and mutant seedlings [13].
 

Analytical, diagnostic and therapeutic context of NIT1

References

  1. Structural analysis of the nit2/nit1/nit3 gene cluster encoding nitrilases, enzymes catalyzing the terminal activation step in indole-acetic acid biosynthesis in Arabidopsis thaliana. Hillebrand, H., Bartling, D., Weiler, E.W. Plant Mol. Biol. (1998) [Pubmed]
  2. Cloning and expression of an Arabidopsis nitrilase which can convert indole-3-acetonitrile to the plant hormone, indole-3-acetic acid. Bartling, D., Seedorf, M., Mithöfer, A., Weiler, E.W. Eur. J. Biochem. (1992) [Pubmed]
  3. Characterization and synthetic applications of recombinant AtNIT1 from Arabidopsis thaliana. Wajant, H., Effenberger, F. Eur. J. Biochem. (2002) [Pubmed]
  4. Arabidopsis mutants resistant to the auxin effects of indole-3-acetonitrile are defective in the nitrilase encoded by the NIT1 gene. Normanly, J., Grisafi, P., Fink, G.R., Bartel, B. Plant Cell (1997) [Pubmed]
  5. Differential regulation of an auxin-producing nitrilase gene family in Arabidopsis thaliana. Bartel, B., Fink, G.R. Proc. Natl. Acad. Sci. U.S.A. (1994) [Pubmed]
  6. Molecular characterization of two cloned nitrilases from Arabidopsis thaliana: key enzymes in biosynthesis of the plant hormone indole-3-acetic acid. Bartling, D., Seedorf, M., Schmidt, R.C., Weiler, E.W. Proc. Natl. Acad. Sci. U.S.A. (1994) [Pubmed]
  7. Genes encoding nitrilase-like proteins from tobacco. Dohmoto, M., Tsunoda, H., Isaji, G., Chiba, R., Yamaguchi, K. DNA Res. (2000) [Pubmed]
  8. Structure of the gene encoding nitrilase 1 from Arabidopsis thaliana. Hillebrand, H., Tiemann, B., Hell, R., Bartling, D., Weiler, E.W. Gene (1996) [Pubmed]
  9. Transgenic tobacco plants expressing the Arabidopsis thaliana nitrilase II enzyme. Schmidt, R.C., Müller, A., Hain, R., Bartling, D., Weiler, E.W. Plant J. (1996) [Pubmed]
  10. The Arabidopsis thaliana isogene NIT4 and its orthologs in tobacco encode beta-cyano-L-alanine hydratase/nitrilase. Piotrowski, M., Schönfelder, S., Weiler, E.W. J. Biol. Chem. (2001) [Pubmed]
  11. Occurrence and formation of indole-3-acetamide in Arabidopsis thaliana. Pollmann, S., Müller, A., Piotrowski, M., Weiler, E.W. Planta (2002) [Pubmed]
  12. Diverse range of gene activity during Arabidopsis thaliana leaf senescence includes pathogen-independent induction of defense-related genes. Quirino, B.F., Normanly, J., Amasino, R.M. Plant Mol. Biol. (1999) [Pubmed]
  13. The tu8 mutation of Arabidopsis thaliana encoding a heterochromatin protein 1 homolog causes defects in the induction of secondary metabolite biosynthesis. Bennett, R.N., Wenke, T., Freudenberg, B., Mellon, F.A., Ludwig-Müller, J. Plant biology (Stuttgart, Germany) (2005) [Pubmed]
 
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