EG10094

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Author Information

Camila Camacho

Basic Information

  • ID: EG10094
  • Name: asnS
  • Organism: E. coli
  • Description: Asparagine-tRNA ligase (AsnRS) is a member of the family of aminoacyl-tRNA synthetases, which interpret the genetic code by covalently linking amino acids to their specific tRNA molecules. The reaction is driven by ATP hydrolysis. AsnRS belongs to the Class II aminoacyl tRNA synthetases, which share three regions of homology Eriani90. AsnRS is a dimer in solution Anselme89, Aoki92. An asparagine-tRNA ligase is an enzyme that catalyzes the chemical reaction ATP + L-asparagine + tRNAAsn -> AMP + diphosphate + L-asparaginyl-tRNAAsn. As an Aminoacyl-tRNA synthetase, the enzyme activates an amino acid for translation by forming an aminoacyladenylate intermediate and then linking this activated amino acid to the corresponding tRNA molecule (amino acid-tRNA, aminoacyl-tRNA). In general, a specific aminoacyl-tRNA synthase is available for each amino acid. Aminoacyl-tRNA synthetases attach amino acids to the 3' termini of cognate tRNAs to establish the specificity of protein synthesis Francklyn2. The 3 substrates of this enzyme are ATP, L-asparagine, and tRNA(Asn), whereas its 3 products are AMP, diphosphate, and L-asparaginyl-tRNA(Asn).
  • DNA Length: 1,401 base pairs.
  • DNA sequence:

ATGAGCGTTG TGCCTGTAGC CGACGTACTC CAGGGCCGTG TAGCCGTTGA CAGCGAAGTC ACCGTGCGCG GATGGGTACG TACCCGCCGA GATTCAAAAG CTGGCATCTC CTTCCTCGCC GTTTATGACG GTTCCTGCTT TGATCCTGTA CAGGCTGTCA TCAATAATTC TCTGCCCAAT TACAATGAAG ACGTCCTGCG TCTGACCACC GGCTGCTCGG TCATTGTGAC GGGTAAAGTC GTGGCGTCGC CGGGCCAGGG GCAACAATTT GAAATTCAGG CCAGCAAGGT TGAAGTTGCT GGTTGGGTTG AAGATCCAGA CACTTACCCG ATGGCGGCAA AACGCCACAG CATTGAGTAT CTGCGTGAAG TCGCTCACCT GCGTCCGCGC ACAAACCTGA TTGGTGCCGT CGCGCGCGTT CGCCATACGC TGGCGCAGGC GCTGCATCGC TTCTTTAACG AGCAGGGATT CTTCTGGGTT TCAACGCCAC TGATTACCGC ATCTGATACC GAAGGTGCAG GCGAAATGTT CCGCGTTTCT ACGCTGGATC TGGAAAACCT GCCGCGTAAC GATCAGGGCA AAGTGGATTT CGACAAAGAC TTCTTTGGTA AAGAGTCTTT CCTGACCGTA TCTGGCCAGT TGAACGGCGA AACCTACGCT TGCGCATTGT CCAAAATTTA TACCTTCGGC CCGACTTTCC GTGCTGAAAA CTCCAACACC AGCCGTCACC TGGCGGAATT CTGGATGCTG GAGCCGGAAG TGGCGTTTGC TAACCTGAAC GATATTGCGG GTCTGGCTGA AGCCATGCTG AAATATGTCT TCAAAGCGGT TCTCGAAGAA CGCGCTGACG ACATGAAATT CTTCGCTGAA CGCGTAGATA AAGATGCCGT TTCACGTCTG GAACGCTTCA TTGAAGCCGA TTTTGCGCAG GTGGATTATA CCGACGCAGT GACCATTCTC GAAAACTGCG GCAGGAAGTT TGAAAACCCG GTTTACTGGG GAGTCGATCT CTCTTCTGAG CATGAGCGTT ATCTGGCGGA AGAACACTTT AAAGCACCGG TAGTGGTTAA AAACTATCCG AAAGATATTA AAGCGTTCTA TATGCGCCTT AACGAAGACG GTAAAACCGT TGCGGCTATG GACGTTCTGG CTCCGGGCAT CGGTGAGATC ATTGGTGGCT CCCAGCGTGA AGAACGTCTG GACGTGCTGG ACGAGCGTAT GCTGGAAATG GGCCTGAATA AAGAAGATTA CTGGTGGTAT CGCGATCTGC GTCGCTACGG TACTGTTCCG CATTCAGGTT TCGGTCTTGG TTTTGAACGT CTGATTGCTT ACGTAACTGG CGTGCAAAAC GTACGTGATG TGATTCCGTT CCCACGTACT CCGCGTAACG CCAGCTTCTA A

  • Amino Acid length: 466 amino acids.
  • Amino Acid sequence:

MSVVPVADVL QGRVAVDSEV TVRGWVRTRR DSKAGISFLA VYDGSCFDPV QAVINNSLPN YNEDVLRLTT GCSVIVTGKV VASPGQGQQF EIQASKVEVA GWVEDPDTYP MAAKRHSIEY LREVAHLRPR TNLIGAVARV RHTLAQALHR FFNEQGFFWV STPLITASDT EGAGEMFRVS TLDLENLPRN DQGKVDFDKD FFGKESFLTV SGQLNGETYA CALSKIYTFG PTFRAENSNT SRHLAEFWML EPEVAFANLN DIAGLAEAML KYVFKAVLEE RADDMKFFAE RVDKDAVSRL ERFIEADFAQ VDYTDAVTIL ENCGRKFENP VYWGVDLSSE HERYLAEEHF KAPVVVKNYP KDIKAFYMRL NEDGKTVAAM DVLAPGIGEI IGGSQREERL DVLDERMLEM GLNKEDYWWY RDLRRYGTVP HSGFGLGFER LIAYVTGVQN VRDVIPFPRT PRNASF

Function and Homologs

  • Product: Asparagine- tRNA ligase
  • Closest homologous proteins: The top (max three) homologous proteins to this protein, as identified by BLAST searches.
    • Name: MULTISPECIES: asparagine--tRNA ligase [Proteobacteria]/ Max score: 964/Query Cover: 100%/E-value: 0.0/Ident: 100%/Accession: WP_000117881.1
    • Name: MULTISPECIES: asparagine--tRNA ligase [Escherichia]/Max score: 963/Query Cover: 100%/E-value: 0.0/Ident: 99%/Accession: WP_000117877.1
    • Name: asparagine--tRNA ligase [Shigella boydii]/Max score: 962/Query Cover: 100%/E-value: 0.0/Ident: 99%/Accession: WP_073691882.1

Expression

  • Expression Level: [High].
  • Expression Level Hypothesis: The asnS gene encodes the Asparagine-tRNA ligase. Asparagine-tRNA ligase catalyzes the attachment of asparagine to tRNA, meaning it helps in translation of the protein. The dataset shows that the expression level for asnS is high. Ligases that attach amino acids to tRNA should be expressed at relatively high abundance since they are an important part of protein synthesis which is an important part of cell life.
  • Expression Level References and Description: E. coli proteome data: File:EcoliProteomicExpressionData.xlsx
  • Expression Time: [Early].
  • Expression Level Hypothesis: asnS activates an amino acid for translation by forming an aminoacyladenylate intermediate and then linking this activated amino acid to the corresponding tRNA molecule. trna synthetases are key to translation and can play a role in editing incorrectly charged trna molecules. Thus, this gene should be required early, but not necessarily at the beginning, as it is not necessary for transcription but is necessary to prepare the proteins for translation.
  • Expression Time References and Description: Aminoacyl-tRNA synthetase

Gene Context

  • Other Components in the functional module: Because this functional module is the enzyme itself, there is no other componen
  • Possible Dependencies: rpsJ binds tRNA to the ribosome, which is an important part of the translation process. This is necessary for the Asparagine-tRNA to be useful.
  • Process: The enzyme activates an amino acid for translation by forming an aminoacyladenylate intermediate and then linking this activated amino acid to the corresponding tRNA molecule (amino acid-tRNA, aminoacyl-tRNA)
    • Inputs: ATP, L-asparagine, tRNAAsn
    • Outputs: AMP, diphosphate, L-asparaginyl-tRNAAsn

Construct

We will handle this - not part of your assignment

  • Synthesis Score: The synthesis score: 1, 2,3
  • Predicted Translation Rate: Prediction of construct translation rate from the RBS calculator
  • Design Notes and Details:
  • GenBank File: A link to the GenBank file. file