Agrobacterium tumefaciens is a bacterium that can infect wound sites in many dif

Question

Agrobacterium tumefaciens is a bacterium that can infect wound sites in many different plants, causing a disease called Crown Gall in which the bacterium genetically ‘reprograms’ plant cells such that they lose their normal control mechanisms and grow into a mass that is basically a tumor. Plant cells in the gall tumor produce large amounts of molecules called opines that serve as nutrients for the A. tumefaciens, providing it with sources of carbon and nitrogen and phosphate.

Another bacterium, Agrobacterium radiobacter K84, produces and secretes a molecule that can kill A. tumefaciens. This antibiotic molecule is called agrocin 84, and it is an attractive approach for killing A. tumefaciens because it has great specificity (i.e., it doesn’t affect bacteria other than tumergenic Agrobacterium species). Unfortunately, there are some strains of A. tumefaciens that are not affected by agrocin 84.

Two important pieces of information about any possible antibiotic are: its molecular target and possible resistance mechanisms. Researchers have investigated these topics for agrocin 84 by asking:

How does it kill A. tumefaciens? What is its target in the A. tumefaciens cell?

How do agrocin-resistant strains of A. tumefaciens differ from the sensitive strains?

Your PAK2.1 questions focus on results relevant to these questions. The questions are on the following page. These same questions are in the PAK2.1 ELMS quiz, and that is where you should submit your answers.

PAK2.2A The target for agrocin 84 in A. tumefaciens

Researchers suspected that agrocin 84 could bind to a cytoplasmic enzyme called LeuS in A. tumefaciens. LeuS catalyzes this reaction:

                  Leucine + tRNALeu + ATP ---------> Leucyl-tRNALeu + AMP + PPi

The researchers purified LeuS from cells in an A. tumefaciens culture and assayed its activity in the absence and presence of agrocin 84. Their results are presented in the graph below.

What is the effect of agrocin 84 on the activity of LeuS?

In an A. tumefaciens cell, what key process would be affected and why would this effect have a negative effect on the viability of A. tumefaciens?

PAK2.2B An agromycin-resistant strain (Mutant 1)

Researchers sequenced the whole genome of an agromycin-resistant strain of A. tumefaciens and compared it to the genome sequence of a sensitive strain. The only difference they found was a missense mutation in the chromosomal gene encoding LeuS.

Explain how a mutation affecting this enzyme could confer resistance.

PAK2.2C Another resistant strain (Mutant 2)

Researchers identified a second agromycin-resistant strain of A. tumefaciens and determined that the mutation was in a gene located on the Ti plasmid. Rather than sequencing this large plasmid (205,000 base pairs!) they first mapped the location of the mutation to a 5,200 base pair region of the plasmid. They then compared the nucleotide sequence of this segment of the mutant plasmid to that of the corresponding region from the wild-type plasmid. Those sequences are in the sequence file on ELMS.

Before attempting to interpret the effect of the mutation, it is useful to determine how many proteins this 5,200 bp segment likely encodes and which one(s) might be affected by the mutation. A starting point for doing this is to determine how many ORFs (open reading frames) are in the sequence. To answer the questions below you will need to use an ORF finder such as https://www.ncbi.nlm.nih.gov/orffinder/

2.2C.1.     How may ORFs of greater than 300 bp are there in the DNA sequences found in the wild-type sequence?

2.2C.2. Each ORF has the potential to encode a protein, but in practice some of the ORFs identified by your search will NOT. What will determine whether any particular ORF is actually a gene that gets transcribed and translated?

2.2C.3. Some of the ORFs identified by orffinder are labeled as having Frame 1, others as Frame 2, and still others as having Frame 3. What do these numbers (1, 2, 3) mean?

2.2C.4. Some of the ORFs identified by orffinder are labeled as Strand + while others are designated as Strand -. What do these labels (+/-) mean?

2.2C.5. Comparing the sequences of the mutant and wild type Ti plasmid segments indicates that there is only one difference: the mutant has a single nucleotide deletion at position 994:

980 1000

       Wild type     TGGGCACCGCGGCGATAGCGG

       Mutant 2     TGGGCACCGCGGCG-TAGCGG

                  Would the mutation identified by the researchers affect an ORF? If so, which one(s) and what would be the expected effect on the protein encoded by this ORF?

2.2C.6. What do you think is the function of the protein encoded by the ORF that you identified in 2.2C.5? Briefly explain your answer.

                  Here’s a suggestion that might help you: try using BLASTP (https://blast.ncbi.nlm.nih.gov)

2.2C.7. Explain how the mutation in Mutant 2 could confer agromycin resistance to A. tumefaciens.

                  Here’s a clue that might help you: Agromycin 84 has been referred to as a ‘Trojan Horse toxin’.

Here are the sequences needed:

>wild type sequence (5.2 kb pTi segment)

GTTTGTAATGTATTGTTGCGTTGCAACAAAACTGTCACCAAACTTTCATTTGACGAAGAACGATTACGCTCATAGTATGAGCGATATAGACCATTTCTTGTTGGAAGGGAAGCGATGGTGTTCAACTCAACTCAGGACCGCCAGGCGAAAATCGTTGAGCTTTTGCGCGATGAGCAGTTTTTGGCTATCGGCAGATTGACGGAGCACTTCCAGATTTCGGTGGCCACTGCGAGGCGCGACTTGAGCGAACTCCACGAGGCCGGACTCTTGCGTCGAACTCACGGCGGTGCGGTCAGCGTCACGCAGGTTACACAGGACAAGCCGAACGCTGCCCGCGCCGTCTGGAATCGGGCGGAGAAGGCGGCCATAGCCGGAGTCGTTGCCGGTATGATCGTCGAGGGTGACACGGTTCTTCTGGACGCCGGTACAACTGCGCTGGAAGTTGCCAAGAAGCTCGCTGACCGCAGAAACCTCACCTTCATCTCGAATGGTCTCGACATTGTCGAGGAATTGACGAGAGGGGAGGGCAAAAGCATCTATTCGGTCGGGGGCGAGTATACCGAAACGAACCGCTCCTTCCGCGGCCCTTTGGCGGAGCAGTTTATCCGCCAGTTCAATGTCGACAAACTCATTCTCAACGCGGCGTCGATCGATGTTGATCGCGGATTGATCTGTACGTCGTCGCCCGTGAACGCGAGTGTCGCTCGCGCCATGATCGAAGTTTCGAGCCGCGTGATCGTCGTCGCGGATCATTCGAAATTCACAAAATCGAGCCTCTCGGTGACGGCCAGGATCGAGGATGTCGGCGTGATCGTCACCGACTCTGGAACCCGAACCATCATTGAGACAATACCGGAAAAGCTGCGGAAGAAGTTCGTTGTTGCGAACTAGGTTCGACGGCCGCGCGCGAATTAAAGCCTCATTCAACAGAGCTGGGAGGAAAACCACATGCTTAAGACCAATCGCAGGAATTTTATGATGGGCACCGCGGCGATAGCGGTCGCTTCAACTGCCGGTGCGAAGTTCACCTTCGCGCAAGAACGCCGGGCGCTTCGCCTGGGTGTGAACGGATTGCCGAACTCGCTGGAGCCGGTCAATGCAATCAGCAATGTCGGCCCGCGTATCGTCAACCAGATATTCGACACGCTGATAGCGCGCGATTTTTTCGCGAAGGGAGCTCCCGGCAATGCCATCGACCTCGTTCCGGCACTTGCCGAGAGCTGGGAGCGCATCGATGAGAAATCGGTTCGCTTCAAGCTGCGGCAGAAGGTCATGTTCCACGATGGTGTTGAACTGACCGCGGACGATGTCGCATACACCTTTTCTTCCGAACGCCTTTGGGGTCCCGAAGCGATCAAGAAAATCCCGCTTGGGAAATCCTATTCGCTTGATTTCGATGAGCCCGTGGTCGAGGACAAATACACAGTCACACTTCGCACAAAGACCCCGAGCTATCTCATCGAGACCTTCGTCGCATCCTGGATGAGCCGCATTGTTCCCAAAGAATACTACAAGAAACTGGGAGCAGTAGATTTCGGTAACAAGCCCGTTGGAACAGGCCCGTACAAGTTCGTGGAATTTGTTGCCGGCGACCGTGTGGTGCTGGAAGCCAACGACGCCTACTGGGGTCCAAAGCCGACTGCATCGAAGATCACCTACCAGATCGTCGCAGAGCCAGCCACACGCGTCGCGGGTCTGATCAGCGGTGAATACGATATCATCACTACCCTGACGCCGGACGACATTCAGCTGATAAATAGCTATCCAGACCTCGAAACGCGCGGGACGCTCATCGAAAACTTCCACATGTTCACCTTCAATATGAACCAAGAAGTTTTCAAGGATAAGAAGCTTCGGCGCGCTCTTGCTCTGGCTGTCAACCGGCCAATCATGGTTGAAGCGCTCTGGAAGAAACAAGCATCTATCCCGGCCGGCTTCAATTTCCCGAACTATGGGGAAACCTTCGATCCAAAGCGTAAGGCCATGGAATATAATGTCGAGGAGGCCAAGCGTCTCGTCAAGGAGAGCGGCTACGATGGAACGCCGATCACCTATCACACGATGGGCAACTATTATGCCAACGCCATGCCTGCGTTGATGATGATGATCGAAATGTGGAAGCAGATCGGTGTGAACGTGGTCATGAAAACCTACGCACCGGGTAGCTTCCCCCCGGACAACCAGACCTGGATGAGAAACTGGTCCAACGGCCAATGGATGACCGATGCCTACGCCACGATCGTCCCCGAATTCGGGCCGAATGGTCAGGTTCAAAAGCGTTGGGGTTGGAAGGCGCCGGCCGAGTTCAACGAATTATGTCAGAAGGTCACGGTTCTGCCGAATGGCAAGGAGCGCTTCGACGCCTACAATCGCATGCGGGACATTTTCGAAGAGGAGGCGCCAGCCGTCATTCTCTATCAGCCGTATGACGTTTATGCCGCGCGCAAGGATGTCCACTGGAAGCCCGTGAGCTTCGAGATGATGGAATTCCGCAACAATCTCAGCTTCGGCTGAACTCGTTAATCGAGAAGGCGGCGCGCGCGAGATATGCGCGTCGCCCTGGAGGAGCCTAAGTTGTCAAATCTACTAACAGTTCGCGACCTGATCGTTCAGGTGCCGGCGCGCGATATCACAATCATCAACGGTGTCAGTTTTTCACTGGATGCAGGGCAGACGCTTGGACTCGTCGGGGAGTCGGGCTGCGGAAAGAGCATGACATGTTACGCAATAGCCAAGGCATTGCCGCGAGGGATTTCGCAGACGGGCGGAACTATTGAGCTGGACAGTGGACCAAAGGCTGATGGCAAGCCGGCTATTGCCATGATCTATCAGGACCCGACCAGCAGCCTCAATCCGGTTCATTCGATCGGTTATTATCTCGAGAGCAGCCTTTATCGCCATCAGGGTCTGGAGGGCAACGATGCACGGCTCGAGGCCATGCGTCTTCTCGAACGTGTCGGTATCGACCGCGCCAAAAGTCGGCTGCGATCCTATCCTCATGAGTTCTCGGGCGGTATGAACCAGCGCGTCATGATTGCCCACGCGCTCGCTGCGAAGCCCAAGCTGATGATCGCCGACGAGCCCACGACAGCGCTCGATGTCACGACGCAGGCGCAGATCCTTCATCTTCTGGAGGAACTCAGATCTGAAACAGGCATGGCGCTTTTGATCGTGTCGCACGATCTCGGCGTGATCGCCCGCCTCGCGGATCGTGCGGCGGTCATGTATTGCGGAAAGATCGTCGAAACGGCGCCGGTTGCGGAATTGTTGGAGCGTGCAGCGCATCCTTACGCTCGCGCCCTGATCGGCTGCATGCCAACGATCGATGCGGACGACCTGGAGCCGCCAGTACCAATCCCGGGTTCTGTTCCTCTTCTCGATAACCTGCCTGCCGGATGTTATTATCACCCACGCTGTCCCCGCGCGAGCGAACAATGTTCCGTGAGCTTTCCCGGGCCTGTCAACGTCGGCGCGTTCCATGACGCGGCCTGTTACCATCCGGTGAGCCCATGAGTGCGCCTCTTCTTCAGGCCCGCGGGGTCACCAAGGTCTTTCGCCTGAAATCCGGGCTTTTCGTAAAGCCATCGGTACATGTGGCGGTTAATTCAATTGATCTCGAACTTGCGCCGCGTGAGCGGGTAGGTGTCGTCGGTGAATCTGGAAGCGGGAAATCGACACTCGGTAGACTTCTGCTCGGCTTGACCTCCACCACCGAGGGAGAGGTCTTGTTCGAAGGACTCGCGCACACAGATCGTAGCGCGCGAGACTGGTCGACGTTCCGAAAGGAGACCTCCCTGGTTCAGCAAAACCCGCTGTCCGCCCTTAATCCACAAATGACGATCGGCGAGCAAATCGCTGAAGCCGTCTGGGTTCACCGGGTTGCCAACCGCGCGGGCGCGCGTGATCGGGCGGCAACCATGCTGGACCGGGTCGGCCTCTCGAGCGCGATGATGAACCGCTATCCACATCAGATGTCGGGAGGGCAGCGACAAAGAGTTGTTATTGCCCGAGCTTTGATCCTCGACCCCAAATTGGTCGTATTCGATGAGGCGGTATCTGCGCTCGACGTTTCTGTGCAAGCCCAGGTCGTTGCGTTGCTTCGGCAGCTCTGGCAGGAGCTTGATCTGGCGTATGTGTTCATCACCCACGACCTGCGCATCGTTCGCCATCTCGTTGATCGTATTGTCGTGATGTATCTTGGCCGCGTCGTCGAAACCGGCGATATCAAATCTGTCTATGCGTGGCCACGCCATCCCTATACCCGCGCATTGCTTGCCTCCGTCCCCACTATGGACCCGACGGTCCGCAATATTGAACCACCGATCAAGGGGGAACTCGACGCTGGCAAGGTAATGACTGGCTGTGCCTTCCGGTCACGCTGTCCTTTTGCAATCGAAAGATGTGCGAAAGAAACGCCGCTGCTTCGGCCTGCGGGCGGGCAACTGGCAGCGTGCCATAGAGCGGAAGAGTTTCCGAGATCTATAGTCGCGCAGACTGATCCGGCGCAGATGATGGAGGTGGCGCGATGATTAGTTTCATCCTTGCACGCCTCGTTCGAGCGATCATTGTCATGATGCTGACCGTGACGTTTGTCTTCATCATTCTCAGGTTGGGTGGAGATCCAGCTCGGGCCATGTTGGGCGAGAACGCGACGCCAGAGGCGCTGGCAGCATTCCGAACAGCATGGGGCCTGGACAGATCCATTCCTGAACAGTTCCTGATCTATCTCGCCAGCGCGCTCAGAGGCGACTTCGGGGTATCGGCAGCCGACGGTCGTGAGGTATTCACCGTCATCTCCGAACGTATACCCAAGACATTGACGCTGACGATCTCAGCATTCGTGCTAAGCATACTCGTCGGTATCCCCGCGGGAATCATCGCAGCAGTCCGCCGCGACAGTGCCGCCGACAAATCCGTCATGGCAGGCGCTGTTCTCGGCTATAGCGTACCAAATTTCTTGCTCGGACTGACGCTCATCTTCGTGTTTGCCGTCTGGTTTCGTGTGCTGCCGAGCTCGGGAAGTTCCACCTGGCAGCATGCGATCCTGCCCGTTGCGACATTCGGTCTTGTCAATGCGGCTGCGATTGCGCGCTTCGCGCGTTCCGCGCTGATCGAAGTTCTGGAACAGCCCTATATCGCAGCGGCTCGCGCCGATGGTATTCCCAAGTGGGAAGTGGTTATCCGCCACGCGCTCCCGAACGCGGCGATCCC

>Mutant 2 sequence (5.2 kb pTi segment)

GTTTGTAATGTATTGTTGCGTTGCAACAAAACTGTCACCAAACTTTCATTTGACGAAGAACGATTACGCTCATAGTATGAGCGATATAGACCATTTCTTGTTGGAAGGGAAGCGATGGTGTTCAACTCAACTCAGGACCGCCAGGCGAAAATCGTTGAGCTTTTGCGCGATGAGCAGTTTTTGGCTATCGGCAGATTGACGGAGCACTTCCAGATTTCGGTGGCCACTGCGAGGCGCGACTTGAGCGAACTCCACGAGGCCGGACTCTTGCGTCGAACTCACGGCGGTGCGGTCAGCGTCACGCAGGTTACACAGGACAAGCCGAACGCTGCCCGCGCCGTCTGGAATCGGGCGGAGAAGGCGGCCATAGCCGGAGTCGTTGCCGGTATGATCGTCGAGGGTGACACGGTTCTTCTGGACGCCGGTACAACTGCGCTGGAAGTTGCCAAGAAGCTCGCTGACCGCAGAAACCTCACCTTCATCTCGAATGGTCTCGACATTGTCGAGGAATTGACGAGAGGGGAGGGCAAAAGCATCTATTCGGTCGGGGGCGAGTATACCGAAACGAACCGCTCCTTCCGCGGCCCTTTGGCGGAGCAGTTTATCCGCCAGTTCAATGTCGACAAACTCATTCTCAACGCGGCGTCGATCGATGTTGATCGCGGATTGATCTGTACGTCGTCGCCCGTGAACGCGAGTGTCGCTCGCGCCATGATCGAAGTTTCGAGCCGCGTGATCGTCGTCGCGGATCATTCGAAATTCACAAAATCGAGCCTCTCGGTGACGGCCAGGATCGAGGATGTCGGCGTGATCGTCACCGACTCTGGAACCCGAACCATCATTGAGACAATACCGGAAAAGCTGCGGAAGAAGTTCGTTGTTGCGAACTAGGTTCGACGGCCGCGCGCGAATTAAAGCCTCATTCAACAGAGCTGGGAGGAAAACCACATGCTTAAGACCAATCGCAGGAATTTTATGATGGGCACCGCGGCGTAGCGGTCGCTTCAACTGCCGGTGCGAAGTTCACCTTCGCGCAAGAACGCCGGGCGCTTCGCCTGGGTGTGAACGGATTGCCGAACTCGCTGGAGCCGGTCAATGCAATCAGCAATGTCGGCCCGCGTATCGTCAACCAGATATTCGACACGCTGATAGCGCGCGATTTTTTCGCGAAGGGAGCTCCCGGCAATGCCATCGACCTCGTTCCGGCACTTGCCGAGAGCTGGGAGCGCATCGATGAGAAATCGGTTCGCTTCAAGCTGCGGCAGAAGGTCATGTTCCACGATGGTGTTGAACTGACCGCGGACGATGTCGCATACACCTTTTCTTCCGAACGCCTTTGGGGTCCCGAAGCGATCAAGAAAATCCCGCTTGGGAAATCCTATTCGCTTGATTTCGATGAGCCCGTGGTCGAGGACAAATACACAGTCACACTTCGCACAAAGACCCCGAGCTATCTCATCGAGACCTTCGTCGCATCCTGGATGAGCCGCATTGTTCCCAAAGAATACTACAAGAAACTGGGAGCAGTAGATTTCGGTAACAAGCCCGTTGGAACAGGCCCGTACAAGTTCGTGGAATTTGTTGCCGGCGACCGTGTGGTGCTGGAAGCCAACGACGCCTACTGGGGTCCAAAGCCGACTGCATCGAAGATCACCTACCAGATCGTCGCAGAGCCAGCCACACGCGTCGCGGGTCTGATCAGCGGTGAATACGATATCATCACTACCCTGACGCCGGACGACATTCAGCTGATAAATAGCTATCCAGACCTCGAAACGCGCGGGACGCTCATCGAAAACTTCCACATGTTCACCTTCAATATGAACCAAGAAGTTTTCAAGGATAAGAAGCTTCGGCGCGCTCTTGCTCTGGCTGTCAACCGGCCAATCATGGTTGAAGCGCTCTGGAAGAAACAAGCATCTATCCCGGCCGGCTTCAATTTCCCGAACTATGGGGAAACCTTCGATCCAAAGCGTAAGGCCATGGAATATAATGTCGAGGAGGCCAAGCGTCTCGTCAAGGAGAGCGGCTACGATGGAACGCCGATCACCTATCACACGATGGGCAACTATTATGCCAACGCCATGCCTGCGTTGATGATGATGATCGAAATGTGGAAGCAGATCGGTGTGAACGTGGTCATGAAAACCTACGCACCGGGTAGCTTCCCCCCGGACAACCAGACCTGGATGAGAAACTGGTCCAACGGCCAATGGATGACCGATGCCTACGCCACGATCGTCCCCGAATTCGGGCCGAATGGTCAGGTTCAAAAGCGTTGGGGTTGGAAGGCGCCGGCCGAGTTCAACGAATTATGTCAGAAGGTCACGGTTCTGCCGAATGGCAAGGAGCGCTTCGACGCCTACAATCGCATGCGGGACATTTTCGAAGAGGAGGCGCCAGCCGTCATTCTCTATCAGCCGTATGACGTTTATGCCGCGCGCAAGGATGTCCACTGGAAGCCCGTGAGCTTCGAGATGATGGAATTCCGCAACAATCTCAGCTTCGGCTGAACTCGTTAATCGAGAAGGCGGCGCGCGCGAGATATGCGCGTCGCCCTGGAGGAGCCTAAGTTGTCAAATCTACTAACAGTTCGCGACCTGATCGTTCAGGTGCCGGCGCGCGATATCACAATCATCAACGGTGTCAGTTTTTCACTGGATGCAGGGCAGACGCTTGGACTCGTCGGGGAGTCGGGCTGCGGAAAGAGCATGACATGTTACGCAATAGCCAAGGCATTGCCGCGAGGGATTTCGCAGACGGGCGGAACTATTGAGCTGGACAGTGGACCAAAGGCTGATGGCAAGCCGGCTATTGCCATGATCTATCAGGACCCGACCAGCAGCCTCAATCCGGTTCATTCGATCGGTTATTATCTCGAGAGCAGCCTTTATCGCCATCAGGGTCTGGAGGGCAACGATGCACGGCTCGAGGCCATGCGTCTTCTCGAACGTGTCGGTATCGACCGCGCCAAAAGTCGGCTGCGATCCTATCCTCATGAGTTCTCGGGCGGTATGAACCAGCGCGTCATGATTGCCCACGCGCTCGCTGCGAAGCCCAAGCTGATGATCGCCGACGAGCCCACGACAGCGCTCGATGTCACGACGCAGGCGCAGATCCTTCATCTTCTGGAGGAACTCAGATCTGAAACAGGCATGGCGCTTTTGATCGTGTCGCACGATCTCGGCGTGATCGCCCGCCTCGCGGATCGTGCGGCGGTCATGTATTGCGGAAAGATCGTCGAAACGGCGCCGGTTGCGGAATTGTTGGAGCGTGCAGCGCATCCTTACGCTCGCGCCCTGATCGGCTGCATGCCAACGATCGATGCGGACGACCTGGAGCCGCCAGTACCAATCCCGGGTTCTGTTCCTCTTCTCGATAACCTGCCTGCCGGATGTTATTATCACCCACGCTGTCCCCGCGCGAGCGAACAATGTTCCGTGAGCTTTCCCGGGCCTGTCAACGTCGGCGCGTTCCATGACGCGGCCTGTTACCATCCGGTGAGCCCATGAGTGCGCCTCTTCTTCAGGCCCGCGGGGTCACCAAGGTCTTTCGCCTGAAATCCGGGCTTTTCGTAAAGCCATCGGTACATGTGGCGGTTAATTCAATTGATCTCGAACTTGCGCCGCGTGAGCGGGTAGGTGTCGTCGGTGAATCTGGAAGCGGGAAATCGACACTCGGTAGACTTCTGCTCGGCTTGACCTCCACCACCGAGGGAGAGGTCTTGTTCGAAGGACTCGCGCACACAGATCGTAGCGCGCGAGACTGGTCGACGTTCCGAAAGGAGACCTCCCTGGTTCAGCAAAACCCGCTGTCCGCCCTTAATCCACAAATGACGATCGGCGAGCAAATCGCTGAAGCCGTCTGGGTTCACCGGGTTGCCAACCGCGCGGGCGCGCGTGATCGGGCGGCAACCATGCTGGACCGGGTCGGCCTCTCGAGCGCGATGATGAACCGCTATCCACATCAGATGTCGGGAGGGCAGCGACAAAGAGTTGTTATTGCCCGAGCTTTGATCCTCGACCCCAAATTGGTCGTATTCGATGAGGCGGTATCTGCGCTCGACGTTTCTGTGCAAGCCCAGGTCGTTGCGTTGCTTCGGCAGCTCTGGCAGGAGCTTGATCTGGCGTATGTGTTCATCACCCACGACCTGCGCATCGTTCGCCATCTCGTTGATCGTATTGTCGTGATGTATCTTGGCCGCGTCGTCGAAACCGGCGATATCAAATCTGTCTATGCGTGGCCACGCCATCCCTATACCCGCGCATTGCTTGCCTCCGTCCCCACTATGGACCCGACGGTCCGCAATATTGAACCACCGATCAAGGGGGAACTCGACGCTGGCAAGGTAATGACTGGCTGTGCCTTCCGGTCACGCTGTCCTTTTGCAATCGAAAGATGTGCGAAAGAAACGCCGCTGCTTCGGCCTGCGGGCGGGCAACTGGCAGCGTGCCATAGAGCGGAAGAGTTTCCGAGATCTATAGTCGCGCAGACTGATCCGGCGCAGATGATGGAGGTGGCGCGATGATTAGTTTCATCCTTGCACGCCTCGTTCGAGCGATCATTGTCATGATGCTGACCGTGACGTTTGTCTTCATCATTCTCAGGTTGGGTGGAGATCCAGCTCGGGCCATGTTGGGCGAGAACGCGACGCCAGAGGCGCTGGCAGCATTCCGAACAGCATGGGGCCTGGACAGATCCATTCCTGAACAGTTCCTGATCTATCTCGCCAGCGCGCTCAGAGGCGACTTCGGGGTATCGGCAGCCGACGGTCGTGAGGTATTCACCGTCATCTCCGAACGTATACCCAAGACATTGACGCTGACGATCTCAGCATTCGTGCTAAGCATACTCGTCGGTATCCCCGCGGGAATCATCGCAGCAGTCCGCCGCGACAGTGCCGCCGACAAATCCGTCATGGCAGGCGCTGTTCTCGGCTATAGCGTACCAAATTTCTTGCTCGGACTGACGCTCATCTTCGTGTTTGCCGTCTGGTTTCGTGTGCTGCCGAGCTCGGGAAGTTCCACCTGGCAGCATGCGATCCTGCCCGTTGCGACATTCGGTCTTGTCAATGCGGCTGCGATTGCGCGCTTCGCGCGTTCCGCGCTGATCGAAGTTCTGGAACAGCCCTATATCGCAGCGGCTCGCGCCGATGGTATTCCCAAGTGGGAAGTGGTTATCCGCCACGCGCTCCCGAACGCGGCGATCCC

Explanation / Answer

Agrocin 84 inhibit the action of LeuS activity by looking like a stable form of Leu-AMP and hence, inhibit the binding of the Leu and ATP substrates.

Agrocin 84 inhibits the nucleic acid and protein synthesis pathway in Agrobacterium tumefaciens. As, inhibition of these pathways, bacterium will not be able to replicate, nor able to make proteins that are required for the viability.

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