EG10034

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Author Information

Basic Information

  • ID: EG10034
  • Name: alaRS
  • Organism: E. Coli
  • Description:This gene is responsible for Alanine--tRNA ligase, which is important for Transfer RNA. [1] RNA comes in three different forms, messenger RNA, transfer RNA, and ribosomal RNA. Transfer RNA (or tRNA) is responsible for desiphering the mRNA, which copies the genetic information from the DNA. [2]

This enzyme is involved in catatylizing the chemical reaction:

ATP + L-alanine + tRNAAla ← → AMP + diphosphate + L-alanyl-tRNAAla

Specifically, this catalyzes the attachment of alanine to tRNA. This occurs in a two-step reaction, first Ala-AMP is formed by the activation of alanine by ATP. Next, Ala-AMP is transferred to the accepter end of tRNA.[3]


  • DNA Length: 2628 base pairs.
  • DNA sequence:

ATGAGCAAATCTACGGCAGAGATCCGCCAGGCATTCCTGGATTTCTTCCACTCAAAGGGGCATCAAGTTGTTGCAAGCTCGTCGTTAGTACCTCACAACGACCCGACTCTTCTGTTCACTAATGCCGGCATGAACCAATTCAAAGATGTGTTTCTGGGTTTGGACAAGCGTAATTATTCGCGTGCCACTACCTCTCAACGCTGCGTGCGCGCTGGAGGTAAACATAATGATTTGGAAAATGTTGGCTATACCGCCCGCCATCACACGTTTTTCGAAATGTTGGGCAACTTCTCTTTTGGTGACTACTTCAAACATGACGCAATTCAATTCGCGTGGGAATTGCTTACGTCAGAGAAGTGGTTTGCCTTGCCAAAAGAGCGTCTTTGGGTCACAGTCTATGAGTCCGACGATGAGGCTTACGAAATCTGGGAGAAAGAGGTAGGTATCCCACGTGAACGTATCATCCGCATCGGCGACAATAAAGGGGCCCCCTACGCAAGTGATAATTTTTGGCAAATGGGTGATACTGGCCCGTGTGGTCCCTGCACCGAAATCTTCTACGACCATGGTGACCATATCTGGGGTGGACCACCTGGATCTCCGGAAGAGGACGGAGATCGCTATATTGAAATTTGGAATATTGTATTCATGCAGTTCAACCGTCAAGCCGATGGTACCATGGAGCCCTTACCCAAGCCTTCCGTGGATACGGGTATGGGGTTAGAACGTATCGCGGCTGTTTTACAGCACGTAAACAGTAATTATGACATTGACTTATTTCGTACATTGATTCAGGCAGTCGCTAAAGTTACGGGTGCCACCGACCTGAGCAATAAATCACTTCGCGTGATTGCAGATCATATCCGTAGTTGCGCTTTTTTAATTGCAGACGGTGTTATGCCTAGTAACGAAAACCGCGGGTATGTACTTCGTCGCATTATCCGCCGCGCAGTGCGCCACGGTAATATGCTTGGGGCAAAAGAAACATTCTTTTACAAATTGGTGGGTCCCCTTATTGATGTCATGGGATCCGCGGGGGAAGATCTTAAGCGTCAACAGGCCCAAGTTGAGCAGGTTTTGAAAACAGAGGAAGAGCAATTTGCACGTACCTTAGAGCGCGGTTTAGCACTGCTGGACGAGGAACTGGCGAAACTTAGTGGAGACACTTTGGACGGAGAGACGGCGTTCCGCTTGTATGATACTTACGGTTTCCCTGTGGACCTTACCGCCGATGTCTGTCGCGAGCGTAATATCAAAGTAGATGAGGCGGGCTTTGAGGCGGCAATGGAGGAGCAGCGTCGTCGCGCACGTGAAGCTTCTGGGTTCGGAGCCGATTATAATGCCATGATTCGCGTCGACTCAGCTTCGGAGTTTAAAGGTTACGACCACTTAGAGCTTAATGGCAAAGTGACAGCGCTTTTTGTGGACGGGAAGGCAGTGGATGCTATCAATGCCGGGCAAGAAGCTGTGGTAGTCCTGGATCAAACACCGTTTTACGCCGAATCTGGTGGGCAGGTAGGGGATAAAGGTGAACTGAAGGGTGCCAACTTTAGTTTCGCAGTGGAGGATACACAGAAATATGGTCAGGCGATTGGACACATTGGAAAATTGGCGGCTGGTAGCTTAAAGGTTGGGGACGCTGTCCAGGCAGACGTCGATGAAGCGCGTCGTGCCCGTATCCGTTTGAATCATTCCGCAACGCATCTTATGCACGCGGCTTTACGTCAAGTGCTTGGCACGCACGTCTCGCAAAAAGGGTCATTGGTGAACGACAAAGTTCTGCGTTTCGATTTTAGTCATAACGAAGCTATGAAACCCGAGGAGATCCGCGCAGTCGAAGATCTGGTTAACACCCAGATTCGCCGTAACTTGCCTATTGAGACAAATATCATGGACCTGGAAGCGGCGAAGGCGAAGGGTGCGATGGCTCTTTTCGGGGAGAAATACGATGAACGCGTTCGCGTCTTGTCGATGGGAGACTTTTCCACAGAGCTTTGTGGTGGCACCCATGCATCGCGCACTGGCGACATCGGTCTGTTTCGTATTATCAGTGAGAGCGGCACAGCCGCCGGGGTCCGCCGCATTGAAGCAGTGACCGGAGAAGGGGCGATCGCAACCGTACACGCCGATTCTGACCGTCTGTCCGAGGTAGCCCATTTGTTGAAGGGAGACTCGAACAATCTGGCAGACAAGGTCCGCAGTGTATTGGAACGCACCCGTCAGTTAGAAAAAGAGCTTCAACAGTTAAAAGAACAGGCGGCCGCTCAAGAATCGGCAAACTTATCCTCGAAGGCGATCGACGTTAACGGTGTCAAGTTATTGGTTTCGGAATTATCCGGTGTTGAACCCAAAATGTTGCGTACAATGGTGGATGATTTAAAGAACCAGTTAGGGTCGACAATTATTGTGCTTGCAACGGTGGTTGAGGGGAAAGTATCGTTAATTGCTGGCGTATCCAAGGATGTAACAGACCGCGTTAAAGCAGGTGAACTTATTGGCATGGTTGCCCAGCAAGTCGGCGGTAAAGGGGGGGGACGCCCCGATATGGCCCAGGCCGGGGGGACGGATGCCGCGGCTCTGCCGGCCGCGCTTGCTAGCGTAAAAGGGTGGGTTTCGGCGAAGCTGCAG

  • Amino Acid length: 876 amino acids.
  • Amino Acid sequence:

MSKSTAEIRQAFLDFFHSKGHQVVASSSLVPHNDPTLLFTNAGMNQFKDVFLGLDKRNYS RATTSQRCVRAGGKHNDLENVGYTARHHTFFEMLGNFSFGDYFKHDAIQFAWELLTSEKW FALPKERLWVTVYESDDEAYEIWEKEVGIPRERIIRIGDNKGAPYASDNFWQMGDTGPCG PCTEIFYDHGDHIWGGPPGSPEEDGDRYIEIWNIVFMQFNRQADGTMEPLPKPSVDTGMG LERIAAVLQHVNSNYDIDLFRTLIQAVAKVTGATDLSNKSLRVIADHIRSCAFLIADGVM PSNENRGYVLRRIIRRAVRHGNMLGAKETFFYKLVGPLIDVMGSAGEDLKRQQAQVEQVL KTEEEQFARTLERGLALLDEELAKLSGDTLDGETAFRLYDTYGFPVDLTADVCRERNIKV DEAGFEAAMEEQRRRAREASGFGADYNAMIRVDSASEFKGYDHLELNGKVTALFVDGKAV DAINAGQEAVVVLDQTPFYAESGGQVGDKGELKGANFSFAVEDTQKYGQAIGHIGKLAAG SLKVGDAVQADVDEARRARIRLNHSATHLMHAALRQVLGTHVSQKGSLVNDKVLRFDFSH NEAMKPEEIRAVEDLVNTQIRRNLPIETNIMDLEAAKAKGAMALFGEKYDERVRVLSMGD FSTELCGGTHASRTGDIGLFRIISESGTAAGVRRIEAVTGEGAIATVHADSDRLSEVAHL LKGDSNNLADKVRSVLERTRQLEKELQQLKEQAAAQESANLSSKAIDVNGVKLLVSELSG VEPKMLRTMVDDLKNQLGSTIIVLATVVEGKVSLIAGVSKDVTDRVKAGELIGMVAQQVG GKGGGRPDMAQAGGTDAAALPAALASVKGWVSAKLQ

Function and Homologs

  • Product: alanine-tRNA ligase
  • Module: ATP binding
  • Closest homologous proteins: The top (max three) homologous proteins to this protein, as identified by BLAST searches.
    • MULTISPECIES: alanine--tRNA ligase [Proteobacteria], 1805 / 100% / 0.0 / 100% link Accession
    • MULTISPECIES: alanine--tRNA ligase [Enterobacteriaceae], 1803 / 100% / 0.0 / 99% link Accession
    • alanyl-tRNA synthetase [Escherichia coli N40607], 1802 / 100% / 0.0 / 99%, link Accession

Expression

  • Expression Level: High
  • Expression Level Hypothesis: This is an important component in the attachment of Alanine to tRNA, which is crucial in translation, a necessary component of function and growth. It would be expected that its expression level would be high.
  • Expression Level References and Description: This level was taken directly from the database for the proteomic data on E. coli expression analysis: [1]
  • Expression Time: Early
  • Expression Time Hypothesis: This protein is crucial for translation, specifically in the creation of tRNA, so it must be expressed early given that it is crucial for translation.
  • Expression Time References and Description: Information was gathered mainly from previous sources, see description.

Gene Context

  • Other Components in the functional module: EG10947 serS, catalyzes the attachment of serine to tRNA [2]
  • Possible Dependencies: EG10098 atpA, produces ATP from ADP, ATP is necessary for the reaction to take place. [3]
  • Process: catalyzes the attachment of alanine to tRNA(Ala)
    • Input: ATP, L-alanine, tRNA(Ala)
    • Output: AMP, diphosphate, L-alanyl-tRNA(Ala)
    • Resource: Uniprot

Construct

  • Synthesis Score: The synthesis score of your construct: 1, 2,3
  • Predicted Translation Rate: Prediction of construct translation rate from the RBS calculator
  • Design Notes and Details: For example, had to use a rare codon to fix folding energy;
  • GenBank File: A link to the GenBank file. file

References

  1. European Bioinformatics InstituteProtein Information ResourceSIB Swiss Institute of Bioinformatics. "Alanine--tRNA Ligase." AlaS - Alanine--tRNA Ligase - Mycoplasma Mycoides Subsp. Mycoides SC (strain PG1) - AlaS Gene & Protein. N.p., 15 Feb. 2017. Web. 16 May 2017.
  2. Lodish, Harvey. "The Three Roles of RNA in Protein Synthesis." Molecular Cell Biology. 4th Edition. U.S. National Library of Medicine, 01 Jan. 1970. Web. 16 May 2017.
  3. "Alanine—tRNA Ligase." Wikipedia. Wikipedia Foundation, n.d. Web. 12 May 2017.