EG10407

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Author Information

Michael Morrissey: 06024019

Basic Information

  • ID: EG10407
  • Name: gltX
  • Organism: E. coli
  • Description: Catalyzes the attachment of glutamate to tRNA(Glu) in a two-step reaction: glutamate is first activated by ATP to form Glu-AMP and then transferred to the acceptor end of tRNA(Glu). Phosphorylation of GltX by HipA prevents it from being charged, leading to an increase in uncharged tRNA(Glu). This induces amino acid starvation and the stringent response via RelA/SpoT and increased ppGpp levels, which inhibits replication, transcription, translation and cell wall synthesis, reducing growth and leading to multidrug resistance and persistence. Source: uniprot.org
  • DNA Length: 1,416 base pairs.
  • DNA sequence:

atgAAAATCA AAACTCGCTT CGCGCCAAGC CCAACAGGCT ATCTGCACGT TGGCGGCGCG CGTACTGCTC TTTACTCCTG GCTTTTTGCA CGTAACCACG GCGGTGAGTT CGTGCTGCGT ATTGAAGACA CCGATCTTGA GCGTTCCACG CCGGAAGCTA TCGAAGCCAT TATGGATGGC ATGAACTGGC TGAGCCTGGA GTGGGATGAA GGTCCGTACT ACCAGACCAA ACGTTTTGAT CGCTACAACG CGGTGATCGA TCAGATGCTG GAAGAGGGCA CTGCTTATAA ATGCTATTGC TCTAAAGAGC GCCTGGAAGC GCTGCGCGAA GAGCAAATGG CGAAAGGTGA GAAGCCGCGT TATGACGGTC GCTGCCGCCA CAGCCATGAG CATCATGCTG ATGATGAACC GTGTGTTGTA CGTTTTGCTA ACCCGCAGGA AGGTTCTGTT GTTTTTGACG ATCAGATCCG TGGTCCGATC GAGTTCAGCA ACCAGGAACT GGACGATCTT ATTATCCGCC GTACCGATGG TTCCCCAACC TATAACTTCT GTGTGGTTGT CGATGACTGG GATATGGAAA TCACCCACGT TATCCGTGGC GAAGACCATA TCAACAACAC GCCACGCCAG ATCAACATTC TTAAGGCCCT GAAAGCGCCG GTGCCGGTTT ACGCGCACGT TTCTATGATC AATGGCGATG ACGGTAAAAA ACTGTCCAAA CGTCACGGGG CAGTCAGCGT AATGCAGTAT CGTGATGACG GTTATTTGCC AGAAGCACTG CTGAACTATC TGGTGCGTCT GGGCTGGTCC CACGGCGATC AGGAAATCTT CACTCGTGAA GAGATGATCA AATACTTCAC TTTGAATGCC GTCAGCAAAT CTGCCAGTGC GTTCAACACC GACAAGCTGC TGTGGCTGAA CCATCACTAC ATTAACGCGC TGCCGCCGGA GTATGTTGCT ACTCACTTAC AGTGGCACAT TGAGCAGGAA AATATCGATA CCCGTAACGG CCCGCAGCTG GCTGATCTGG TGAAACTGCT GGGCGAACGC TGCAAGACGC TGAAAGAGAT GGCACAGAGC TGCCGTTATT TCTACGAAGA TTTTGCTGAG TTCGATGCCG ACGCCGCGAA AAAACATCTG CGTCCGGTAG CGCGTCAGCC GCTGGAAGTG GTTCGTGACA AACTGGCCGC GATTACTGAC TGGACCGCTG AAAACGTTCA TCACGCTATT CAGGCGACGG CGGATGAGCT GGAAGTGGGT ATGGGTAAAG TTGGTATGCC GCTGCGTGTC GCCGTAACCG GTGCGGGGCA GTCTCCAGCA CTGGATGTTA CCGTTCACGC AATTGGTAAG ACCCGCAGTA TCGAGCGTAT CAACAAAGCG CTGGATTTTA TTGCTGAACG CGAAAATCAG CAGtaa

  • Amino Acid length: 471 amino acids.
  • Amino Acid sequence:

MKIKTRFAPS PTGYLHVGGA RTALYSWLFA RNHGGEFVLR IEDTDLERST PEAIEAIMDG MNWLSLEWDE GPYYQTKRFD RYNAVIDQML EEGTAYKCYC SKERLEALRE EQMAKGEKPR YDGRCRHSHE HHADDEPCVV RFANPQEGSV VFDDQIRGPI EFSNQELDDL IIRRTDGSPT YNFCVVVDDW DMEITHVIRG EDHINNTPRQ INILKALKAP VPVYAHVSMI NGDDGKKLSK RHGAVSVMQY RDDGYLPEAL LNYLVRLGWS HGDQEIFTRE EMIKYFTLNA VSKSASAFNT DKLLWLNHHY INALPPEYVA THLQWHIEQE NIDTRNGPQL ADLVKLLGER CKTLKEMAQS CRYFYEDFAE FDADAAKKHL RPVARQPLEV VRDKLAAITD WTAENVHHAI QATADELEVG MGKVGMPLRV AVTGAGQSPA LDVTVHAIGK TRSIERINKA LDFIAERENQ Q

Function and Homologs

  • Product: glutamate—tRNA ligase.
  • Closest homologous proteins: The top (max three) homologous proteins to this protein, as identified by BLAST searches.
    • glutamate--tRNA ligase [Shigella sonnei], 988/100%/0.0/99%, [1]
    • glutamate--tRNA ligase [Shigella flexneri], 988/100%/0.0/99%, [2]
    • Glutamyl-tRNA synthetase [Klebsiella pneumoniae IS22], 987/100%/0.0/99%, [3]

Expression

  • Expression Level: High
  • Expression Level Hypothesis: E. Coli may have high levels of proteins that involve glutamate as one or more of their Amino Acids. If this is the case, it makes sense that the cell would need several tRNA binding agents for glutamate to aid in the synthesis of these proteins.
  • Expression Level References and Description: Expression Level was found in the following documentation: Proteomic Expression Data
  • Expression Time: right at the beginning
  • Expression Level Hypothesis: Because this protein is integral in transcribing future proteins that use the amino acid glutamate, it is essential to have right at the begginning to accommodate any other early proteins that require glutamate.
  • Expression Time References and Description: Expression time was estimated based on information on gene function found at []

Gene Context

  • Other Components: HipA, this is an inhibitor that acts in this functional module as an inhibitor, controlling production levels
  • Possible Dependencies: Zn2+, ATP, glutamate, tRNA(Glu)
  • Process: attaches glutamate to the appropriate tRNA
    • Inputs: ATP, L-glutamate, tRNA(Glu)
    • Outputs: AMP, diphosphate, L-glutamyl-tRNA(Glu)

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