MMSYN1 0260

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Author Information

Luísa Galhardo

Basic Information

  • ID: MMSYN1_0260
  • Name: ValS
  • Organism: JCVI-Syn3.0
  • Description:

This protein is an enzyme catalyzing the reaction a tRNAval + L-valine + ATP → an L-valyl-[tRNAval] + AMP + diphosphate, covalently linking L-valine to its specific tRNA molecule using ATP hydrolysis (EcoCyc). Fittingly, the enzyme also binds ATP in addition to Valine. ValRS can inadvertently process amino acids similar to valine, like threonine, so it has a "posttransfer" editing activity that hydrolyzes mischarged Thr-tRNA(Val) in a tRNA-dependent manner (UniProt). ValRS has two distinct active sites: one for aminoacylation and one for editing. The misactivated threonine is translocated from the active site to the editing site (UniProt). This enzyme works with the Ribosome to translate mRNA to an amino acid sequence.

  • DNA Length: 2,619 base pairs.
  • DNA sequence:

ATG AAA AAG CAG TTG AAT CCC AAG TAC GAT CAT TTA CAA GTA GAA AAG GAC AAA TAC CAA TAT TGG TTA GAC AAG AAC CTG TTC AAG GCC GAT ATT AACAGC AAT AAA CCA AAG TTC TCC ATT ATC TTG CCC CCT CCA AAT GTA ACC GGG AAG CTG CAT ATC GGC CAT GCG TGG GAC ACC TCT CTG CAA GAT GCG ATCATC CGC TTC AAA AAA TTA ACG GGG TAT GAT ACG TTG TTT CTG CCA GGC ATG GAC CAT TCC GGT ATC AGC ACT CAG GTG AAA GTA GAA GCT AAG TTA AAGCAA CAA GGC ATC GAT AAG AAT AAG CTG GGC CGT GAA AAA TTT TTA TTA GAG GCC TGG AAG TGG AAG GAA GAA TAT GCA AAC ATT ATT CGT AAA CAA TGGGCT AAA TTA GGT CTG AGT CTG GAC TAT TCG ACT GAG AAG TTC ACA TTG GAT GAG GAT ATC AAC CAG ATC GTC AAG GAG ATT TTT GTT AAA TTT TAC AATGAT CAA ATC ATC TAC AAA GAC AAA CAG ATT GTT AAT TGG GAC CCC GAA CAA AAA ACC GCT ATT TCA AAC GTC GAG GTC ATT TAT AAG GAA ACA CAA TCTAAG ATG TAT TAT TTC AAA TAT ATG TTA GAA GAT TCA AAC GAA TAC TTA ACC GTC GCG ACG ACA CGT CCA GAA ACC ATG TTT GTG GAT CAG TGT TTA GTCGTA AAC CCG AAT GAT TCC CGT TAC CAG AAG TAT ATC AAC AAG AAA GTT ATC AAT CCA GTC AAC AAA CAA GTA ATC CCG ATT ATC AGC GAT GAA TAT GTAGAC ATT GAG TTC GGG ACC GGG GTC ATG AAA TGC ACT CCT GCT CAC GAC TTG AAT GAC TAT CAC TTA GCT ATT AAA CAC AAT TTA AAG ATG CCG ATC TGTCTT AAC ATT GAC GGA AGC GTT AAT CAA TTG GGA GGG AAA TAT CAG GGG CTG GAC CGT TTT GTG GCG CGC AAG GAG ATT ATC AAA AAC GCT ATC AAC GAAGAC TTG TTC GTT AAA GAG GAG GAT ATT ATC AAT CAG GTT GGG TTC TCT GAA CGT AGC AAC GCG ATC GTA GAG CCA TAC CTG TCG GAT CAG TGG TTC GTGAAA ATG GAC AAG TTC AAG AAT ATG GTG ATC GAT CTT CAG AAC AGT GAT AAC AAG ATT AAT TTC TAT CCG AAT CGT TTC TCA GAC GTT CTG AAC CGT TGGATG ACC AAC GCT CAT GAC TGG TGC ATT AGC CGT CAG CTG TGG TGG GGA CAT CAA ATT CCA TGT TGG TAT CAT AAG AAA ACC AAC GAG ATG TAT GTC GGTATC AAC CCG CCG TCG GAT ATT GAA AAC TGG ACC CAG GAT CAG GAT GTA CTG GAT ACC TGG TTT TCC TCT GGC CTG TGG GCT TTC TCT ACT TTG CTG AACAAC AAA GGG TTG GAG TCC GAG TAT TTT AAG AAT TAC TTC CCA ACA TCT GTT CTT GTC ACA GGG TAC GAC ATC ATC TTT TTT TGG GTG GCT CGT ATG ATTTTT CAG ACC TTA GAA TAC ACG AAG CAG ATT CCT TTC AAA GAT GTT CTG ATT CAT GGG CTT GTT CGC GAT GAA CTG AAT CGT AAG ATG AGC AAA TCT CTTGGG AAC GGT ATC GAT CCT ATG GAT GTG ATT AAT AAT AAT GGA TGT GAC AGC CTG CGC CTG TTT CTT TTA ACC AAT AGC ACG CCG GGA CAA GAT ATT CGTTAT TCG AAT GAA AAA ATC CTT GCA AGT TCG AAT TTC ATC AAC AAA CTT TGG AAC GCT TCT CGT TAC GTG TTT TTA AAC TTA GAT AAA GAC TTC AAA TTTGAT CAA AAC TTC TAC AAG ACG GAC TTA GAG ATC ACT AAC CAG TGG ATT TTA ACT GAA TTA TCG AAG ACG CAG GCT TAC GTG TAT GAG AAG ATG AAT AAGTAT GAG TTC AGT CTT GCG GGC AAC CAT TTA TGG GAC TTC GTA TGG AAT AAG TAC TGT AGC TGG TAT ATT GAA TTT AGC AAA GTC AAC TTG AAT AAT GATAAA TTT AAT CAC CAA ACG AAG CAG ACC CTG TTT TAT GTC CTG AAG GAA ATC CTG ATC ATG ATT CAT CCG CTG ATT CCG TTC GTG AGT GAA GAG ATC TACTTG AAC ATG ATG TTG AAG GAA TCA ATC TTG CTG GAG CAG TGG ACA AAC TTA AAC AGT AAC TAT GAC ACC TCG TTT ATC GAC GAT GTT ATC AAG ATG ATCACG AGC ATC CGC GAG TTC AAA AAC ACT AAA AAC ATT AAA AAC GAT GTG TGT CTG TCC GTA AAC GTG TCC AAC ACC AAT CAG AAC CAT ACC AAA CTT TTTAAA AAG CAC TTT GAC CAA ATT TAC TCC TTC CTG ATC AAC TTT TGT AAC ACG AAA TTA GTA GAC TAT GTG ATT AAA AAC AAG ACA AGT CTG TCT ATT GACGAG TAT TTT ATC GAG ATT GCC AAC GAT TCA TTT ATC AAT AAG AAT GAA TTA ATC AAG GAG CTT GAG CAA AAG CAA AAT CAC CTT AAC AAT GAG ATT ACCCGT TCT CAG AAG ATT TTA AAC AAC CAG GAA TTC ATC AAA AAA GCG AAA CCA GAA AAG ATT CAG TCC GAG AAA ATC AAA TAT CAG AAC TAC TTG GAC CAGCTT CAG GCG ATC AAG GAT AAA TTA AAG GAA TTA AAG AAT GAT TAG

  • Amino Acid length: 872 amino acids.
  • Amino Acid sequence:

MKKQLNPKYDHLQVEKDKYQYWLDKNLFKADINSNKPKFSIILPPPNVTGKLHIGHAWDTSLQDAIIRFKKLTGYDTLFLPGMDHSGISTQVKVEAKLKQQGIDKNKLGREKFLLEAWKWKEEYANIIRKQWAKLGLSLDYSTEKFTLDEDINQIVKEIFVKFYNDQIIYKDKQIVNWDPEQKTAISNVEVIYKETQSKMYYFKYMLEDSNEYLTVATTRPETMFVDQCLVVNPNDSRYQKYINKKVINPVNKQVIPIISDEYVDIEFGTGVMKCTPAHDLNDYHLAIKHNLKMPICLNIDGSVNQLGGKYQGLDRFVARKEIIKNAINEDLFVKEEDIINQVGFSERSNAIVEPYLSDQWFVKMDKFKNMVIDLQNSDNKINFYPNRFSDVLNRWMTNAHDWCISRQLWWGHQIPCWYHKKTNEMYVGINPPSDIENWTQDQDVLDTWFSSGLWAFSTLLNNKGLESEYFKNYFPTSVLVTGYDIIFFWVARMIFQTLEYTKQIPFKDVLIHGLVRDELNRKMSKSLGNGIDPMDVINNNGCDSLRLFLLTNSTPGQDIRYSNEKILASSNFINKLWNASRYVFLNLDKDFKFDQNFYKTDLEITNQWILTELSKTQAYVYEKMNKYEFSLAGNHLWDFVWNKYCSWYIEFSKVNLNNDKFNHQTKQTLFYVLKEILIMIHPLIPFVSEEIYLNMMLKESILLEQWTNLNSNYDTSFIDDVIKMITSIREFKNTKNIKNDVCLSVNVSNTNQNHTKLFKKHFDQIYSFLINFCNTKLVDYVIKNKTSLSIDEYFIEIANDSFINKNELIKELEQKQNHLNNEITRSQKILNNQEFIKKAKPEKIQSEKIKYQNYLDQLQAIKDKLKELKND

Function and Homologs

  • Product: Valine—tRNA ligase
  • Closest homologous proteins: The top (max three) homologous proteins to this protein, as identified by BLAST searches.
    • Valine—tRNA ligase [Mesoplasma florum L1], 1119/99%/0.0/63%, Q6F1B1.1
    • Valine—tRNA ligase [Staphylococcus aureus subsp. aureus], 884/99%/0.0/48%, Q6G8R2.1
    • Valine—tRNA ligase [Staphylococcus aureus subsp. aureus], 884/99%/0.0/48%, Q6GG42.1
  • Equivalent E. coli protein: EG11067.

Expression

  • Expression Level: medium
  • Expression Level Hypothesis:

This protein helps in translation, meaning it is probably relatively important to be abundant in the cell, but one protein can continually charge several tRNAs with valine and valine is only 1 of many amino acids, so the expression level may not be the highest.

  • Expression Level References and Description:

The valS protein in File:MgenitaliumSimProteinCounts.xlsx indicates a medium level for this protein.

  • Expression Time: At which time should the gene be expressed in the lifecycle of our organism? early
  • Expression Time References and Description:

According to the Wikipedia article on trna synthetases, these proteins are key to translation and can also play a role in editing incorrectly charged trna molecules. This leads me to believe that it is required early, but not necessarily at the beginning, because it is not necessary for transcription but is necessary to translate proteins.

Gene Context

  • Other Components:

Valine-tRNA ligase does not require any other proteins to perform its function, so there are not really other components. There are however other tRNA ligases that also work to make translation possible, like LysS.

  • Possible Dependencies:

This ligase depends on the presence of valine, for obvious reasons, so the charged trna can be formed.

  • Process: charging tRNA
    • Inputs: tRNAval, L-valine, ATP
    • Outputs: L-valyl-[tRNAval], AMP, diphosphate

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