MMSYN1 0076

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Author Information

Camila Camacho

Basic Information

  • ID: MMSYN1_0076
  • Name: asnRS
  • Organism: JCVI-Syn3.0
  • 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: 1,362 base pairs.
  • DNA sequence:

ATG GAG ATC CGT CAG ATT TTC GAG CAA CAT TCA GAG TTA TTG GAC AAA GAG GTG GAA ATC TTG GGT CGC GTC CGC AGT AAC CGT CAA GGT AAG TTT GTT TCT TTT ATG ATC CTG AAT GAT GGG ACA ACC TTC ACC GAT CTT CAA GTT GTA TAT AAA ACG AAG ACT AAA GGT TAC GAG CAG GCA CTT CAA GCG CGC GTA TCA AGC ATT GTC AAG GTC ATC GGG AGA GTC GTT CTT ACA CCT GAA AAG CAA CAG AAA TTC GAA TTA CAA GCC GAT GAG ATC GAG TTG ATT GAT CAG GCC ATT GAG GAT TAC CCT TTG CAG AAG AAA GAG CAC ACC ACA GAG TAT TTA CGT GAG ATA GCG CAT CTT CGT GCC AAG ACT AAA ACA TTC AAC GCG ATT TTC AAA ATT CGG AGT GCC GCC GCC TAC GCA ATC CAT CGG TTT TTC AAT GAA CGC AAC TTC GTC TAC ATT CAC TCA CCC ATA ATA ACA TCT AAC GAT GCC GAG GGC GCA GGC GAG GCT TTC CTG GTG ACC ACA CGT GAG GAC GCG GAC TAT GAA AAG GAT TTC TTC GGG AAA AAG GCC AGT CTT ACT GTA TCA GGT CAG TTG CAC GCA GAG GCT TTT GCC CAG GCG TTT AAG AAG GTC TAC ACC TTT GGG CCG ACG TTT AGA GCG GAA AAT TCG AAC ACA GCA AAA CAT GCC GCG GAA TTC TGG ATG ATC GAA CCT GAG GTT GCA TTT GCC GAT TTA AAA GAC AAT ATC CAA CTT ATC CAA GAC ATG GTA AAG TAC ATT ATT AAT TAT ATT TTC AAA CAC AAC AGA CGC GAG CTT GAG TTT TGT AAC GAA CAC CTG GAA AAC GGA CTT ATC GAC AAG CTG AAT AAT GTG AGA AAC TCC GAA TTC AAG ATT ACG ACT TAT ACG CAG GCT ATT GAG ATT TTG AAA CAG GCG GTT AAA GAC GGT CAT AAA TTC GAA GTC TCG GAC ATT GAG TTT GGC TTG GAC TTA GGC ACT GAG CAT GAG AGA TAC ATC TGC GAG CAA GTC AAC AAG GCA CCA ACA TTT GTT ACT AAC TAT CCG AAG GAA ATA AAA GCT TTT TAC ATG AAA CAG AAC GAC GAC AAC AAG ACT GTA GCG GCT GTC GAT TTG CTG GTT CCG GGG ATT GGG GAG TTG GTT GGC GGT AGT CAG CGT GAG GAC AAC TAT GAA AAG CTT ATT AAA AGA TGC AAG GAG GTA AAT ATT GAT ATA GAT CAA CTG GAG TGG TAC AAT AAC TTG CGT CTG TAT GGA TAT TAC AAA AGT GCC GGA TTC GGC CTG GGT TTC GAG CGT TTA GTC ATG TAT ATA ACC GGC GCC TCA AAT ATT CGC GAT GTA ATT CCT TTT CCT AGA ACC CCG AAA AAT TTA CTG TTC TAA

  • Amino Acid length: 454 amino acids.
  • Amino Acid sequence:

MEIRQIFEQHSELLDKEVEILGRVRSNRQGKFVSFMILNDGTTFTDLQVVYKTKTKGYEQALQARVSSIVKVIGRVVLTPEKQQKFELQADEIELIDQAIEDYPLQKKEHTTEYLREIAHLRAKTKTFNAIFKIRSAAAYAIHRFFNERNFVYIHSPIITSNDAEGAGEAFLVTTREDADYEKDFFGKKASLTVSGQLHAEAFAQAFKKVYTFGPTFRAENSNTAKHAAEFWMIEPEVAFADLKDNIQLIQDMVKYIINYIFKHNRRELEFCNEHLENGLIDKLNNVRNSEFKITTYTQAIEILKQAVKDGHKFEVSDIEFGLDLGTEHERYICEQVNKAPTFVTNYPKEIKAFYMKQNDDNKTVAAVDLLVPGIGELVGGSQREDNYEKLIKRCKEVNIDIDQLEWYNNLRLYGYYKSAGFGLGFERLVMYITGASNIRDVIPFPRTPKNLLF

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: asparagine--tRNA ligase [Mycoplasma mycoides], Max score: 938/Query Cover: 100%/E-Value: 0.0/Ident: 100%, WP_013729224.1
    • Name: asparagine--tRNA ligase [Mycoplasma leachii], Max score: 915/Query Cover: 100%/E-Value: 0.0/Ident: 97%, WP_013448102.1
    • Name: asparagine--tRNA ligase [Mycoplasma feriruminatoris], Max score: 910/Query Cover: 100%/E-Value: 0.0/Ident: 96%, WP_008362895.1
  • Equivalent E. coli functional protein: EG10094.

Expression

  • Expression Level: [Low].
  • 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 asparagine, so the expression level does not necessarily have to be the highest. The dataset shows that the expression level for asnS (a related gene) is low, meaning that asparagine might not be an amino acid commonly used in this organism's proteins. But, in general, ligases that attach amino acids to tRNA should be expressed at relatively high abundance because they are an important part of protein synthesis which is an important part of cell life.
  • Expression Level References and Description: M. genitalium model data: File:MgenitaliumSimProteinCounts.xlsx
  • Expression Time: [early].
  • Expression Level Hypothesis: asnRS 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 component.
  • 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