EG12401

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Author Information

Jon Bartlett

Basic Information

  • ID: EG12401
  • Name: rlmN
  • Organism: E. coli
  • Description: The gene rlmN codes for a methyltransferase that causes the methylation of 23S rRNA at the C2 position of the A2503 nucleotide.Toh The enzyme has the capability of using the free 23s rRNA as a substrate, but it is incapable of using the fully assembled ribosomal subunit. Additionally, it is responsible for the methylation of certain tRNAs at the C2 position of A37. BenitezPaez The additional requirements for this protein are two molecules of SAM, with one being used as a methyl donor group, and the other as a source of 5'-deoxyadenosyl radical. Yan Cysteine residues are another major requirement for its function.
  • DNA Length: 1155 base pairs.
  • DNA sequence:

ATGAGTGAGCAACTTGTCACTCCAGAGAATGTCACCACAAAGGATGGAAAAATTAACCTTTTGGACCTGAATCGTCAACAAATGCGTGAATTCTTTAAGGATCTGGGCGAGAAACCCTTCCGTGCGGACCAAGTGATGAAGTGGATGTACCATTATTGCTGCGACAATTTCGACGAAATGACAGACATCAACAAAGTGCTGCGTGGAAAACTTAAAGAAGTAGCGGAGATCCGCGCTCCGGAAGTTGTAGAAGAGCAGCGTTCCTCGGACGGTACTATCAAGTGGGCTATTGCAGTCGGGGATCAGCGTGTAGAGACCGTATACATCCCTGAGGATGATCGCGCGACGCTTTGTGTCTCCTCGCAGGTTGGGTGTGCCTTGGAGTGCAAATTTTGCAGTACAGCACAACAAGGCTTTAATCGCAACCTGCGTGTAAGCGAGATTATCGGCCAAGTCTGGCGTGCAGCGAAGATCGTGGGTGCAGCCAAAGTCACCGGACAACGCCCAATTACAAATGTTGTGATGATGGGCATGGGCGAGCCACTGCTTAATCTGAATAACGTAGTTCCAGCGATGGAGATCATGCTTGATGATTTCGGCTTTGGACTTAGTAAACGCCGCGTAACCCTTTCGACCAGTGGAGTAGTTCCTGCATTAGACAAACTTGGGGATATGATTGATGTTGCTTTGGCTATCAGCCTGCATGCGCCTAACGACGAAATTCGTGACGAAATTGTACCGATTAACAAGAAATATAATATCGAGACGTTCCTTGCCGCGGTACGTCGTTATCTGGAAAAAAGTAACGCAAACCAGGGCCGCGTGACTATTGAATACGTGATGTTGGATCATGTTAACGACGGCACCGAGCATGCACACCAGCTGGCAGAGTTATTAAAGGACACTCCCTGCAAGATTAACTTGATCCCTTGGAACCCGTTTCCGGGTGCCCCATATGGACGTTCTTCAAATTCGCGTATTGACCGCTTCTCTAAAGTGCTTATGAGTTACGGCTTCACAACAATTGTTCGTAAAACGCGTGGGGATGACATTGATGCGGCATGCGGCCAGCTGGCGGGCGATGTAATCGATCGTACCAAACGCACATTGCGTAAACGCATGCAAGGTGAAGCTATTGACATTAAAGCGGTGTAA

  • Amino Acid length: 384 amino acids.
  • Amino Acid sequence:

MSEQLVTPENVTTKDGKINLLDLNRQQMREFFKDLGEKPFRADQVMKWMYHYCCDNFDEM TDINKVLRGKLKEVAEIRAPEVVEEQRSSDGTIKWAIAVGDQRVETVYIPEDDRATLCVS SQVGCALECKFCSTAQQGFNRNLRVSEIIGQVWRAAKIVGAAKVTGQRPITNVVMMGMGE PLLNLNNVVPAMEIMLDDFGFGLSKRRVTLSTSGVVPALDKLGDMIDVALAISLHAPNDE IRDEIVPINKKYNIETFLAAVRRYLEKSNANQGRVTIEYVMLDHVNDGTEHAHQLAELLK DTPCKINLIPWNPFPGAPYGRSSNSRIDRFSKVLMSYGFTTIVRKTRGDDIDAACGQLAG DVIDRTKRTLRKRMQGEAIDIKAV

Function and Homologs

  • Product: RNA Methyltransferase
  • Closest homologous proteins: The top (max three) homologous proteins to this protein, as identified by BLAST searches.

Expression

  • Expression Level: Low
  • Expression Level Hypothesis: Methyltransferases in general will have low expression levels because of their efficiency and niche targets. This one specifically targets a specific nucleic acid on a rRNA molecule, thus not many of these proteins need be produced.
  • Expression Level References and Description: Information on the gene and function was obtained here: 1.
  • Expression Time: Late
  • Expression Level Hypothesis: Similar to before, because methyltransferase's main function is to alter an existing structure, their expression will be late because the development of the existing structure must occur first.
  • Expression Time References and Description: This hypothesis was formed by the consideration of the function of the protein and when it is necessary to occur.

Gene Context

  • Other Components in the functional module. rsmG rsmG is a methyltransferase that targets a specific nucleic acid in rRNA, similar to this gene rlmN.
  • Possible Dependencies: An intact, complete Ribosome.
  • Process: rRNA Methylation
    • Inputs: 2 S-adenosyl-L-methionine, adenine(2503) in 23S rRNA, 2 reduced [2Fe-2S] ferredoxin
    • Outputs: S-adenosyl-L-homocysteine, L-methionine, 5'-deoxyadenosine, 2-methyladenine(2503) in 23S rRNA, 2 oxidized [2Fe-2S] ferredoxin

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