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tsaE [2017-09-16 17:16:02]
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tsaE [2017-09-16 17:16:02]

P-loop ATPase, required for threonyl carbamoyl adenosine (t6A) modification of tRNAs that pair with ANN codons in mRNA
locus
BSU05910
pI
4.60
mw
17.76 kDa
protein length
158 aa Sequence Blast
gene length
474 bp Sequence Blast
function
tRNA modification
product
tRNA modification enzyme
essential
no
synonyms
ydiB
Outlinks
KEGGBsubCycSubtiListUniProtExpression Browser

Genomic Context

      

categories

  • [category|SW 3|Information processing] → [category|SW 3.3|Protein synthesis, modification and degradation] → [category|SW 3.3.1|Translation] → [category|SW 3.3.1.8|tRNA modification and maturation]
  • [category|SW 3|Information processing] → [category|SW 3.3|Protein synthesis, modification and degradation] → [category|SW 3.3.4|Protein modification] → [category|SW 3.3.4.2|Protein kinases]
  • [category|SW 4|Lifestyles] → [category|SW 4.3|Coping with stress] → [category|SW 4.3.8|Resistance against oxidative and electrophile stress]

    • Gene

    Coordinates
    641,654 → 642,130

    Phenotypes of a mutant

  • reduced growth rate [Pubmed|19246765]

    • The protein

    Catalyzed reaction/ biological activity

  • biosynthesis of the threonylcarbamoyladenosine (t6A) residue at position 37 of ANN-decoding tRNAs using L-threonylcarbamoyl-AMP ([protein|6CB350C335603D5F4434679F9D740BFECA738599|TsaB]-[protein|30393F16CFE89E7BAD233A0BD27B77D679F7E409|TsaD]-[protein|E15BB518649B2B9CD44CEBBB4FA481E55BCE823C|TsaE]) [Pubmed|23072323,21183954]

    • Protein family

  • UPF0079 family (according to Swiss-Prot)

    • Kinetic information

  • K(M) for ATP: 60 myM, V(max) 10 nmol/min [Pubmed|19246765]

    • Structure

  • [PDB|1HTW] [Pubmed|12112691] (from '' Haemophilus influenzae'', 40% identity)

    • [SW|Localization]

  • cytoplasm with the exception of the nucleoid region [Pubmed|19246765]

    • Additional information

  • ATPase activity is required for ''in vivo'' function of the protein [Pubmed|19246765]

    • Expression and Regulation

    Operons

    genes
    [gene|DE32F1DA1EA1164411E0605262381BD38A49656C|thiL]-[gene|E15BB518649B2B9CD44CEBBB4FA481E55BCE823C|tsaE]-[gene|6CB350C335603D5F4434679F9D740BFECA738599|tsaB]-[gene|C2283BDD806221A285A27F24F52F92F4D47F0C55|ydiD]-[gene|30393F16CFE89E7BAD233A0BD27B77D679F7E409|tsaD]
    description
    [Pubmed|22383849]
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    Biological materials

    Mutant

  • BKE05910 (Δ[gene|E15BB518649B2B9CD44CEBBB4FA481E55BCE823C|tsaE]::erm trpC2), available at [http://www.bgsc.org/getdetail.php?bgscid=BSU05910 BGSC], upstream reverse: 
    _UP1_TTGCTTCACACGAGTCACCC
    , downstream forward: 
    _UP4_ATGCTTTGTGAGGAGTTAAG
  • BKK05910 (Δ[gene|E15BB518649B2B9CD44CEBBB4FA481E55BCE823C|tsaE]::kan trpC2), available at [http://www.bgsc.org/getdetail.php?bgscid=BSU05910 BGSC], upstream reverse: 
    _UP1_TTGCTTCACACGAGTCACCC
    , downstream forward: 
    _UP4_ATGCTTTGTGAGGAGTTAAG

    • Expression vector

  • for expression/ purification from ''B. subtilis'' with N-terminal Strep-tag, for [SW|SPINE], in [SW|pGP380]: pGP833, available in [SW|Stülke] lab
  • for expression/ purification from ''E. coli'' with N-terminal His-tag, in [SW|pWH844]: pGP841, available in [SW|Stülke] lab

    • References

  • 17005971,19246765,23072323,22383849,12112691