Consider the genome of the bacterium Mycoplasma capricolum, which causes several
ID: 90413 • Letter: C
Question
Consider the genome of the bacterium Mycoplasma capricolum, which causes several types of respiratory ailments in humans. There is a strong codon bias in this organism. Explain why this bias exists, and how is would be reflected in the translation of glutamic acid codons. For this amino acid in this draw the complete molecular structure of nucleotide positions #33, 34, 35, and 36 in the tRNA cognate for the codon that is used for this amino acid. Explain how the situation for glutamic acid codon and anticodon usage differs in Mycococcus luteus. Be very explicit.Explanation / Answer
Mycoplasma capricolon :
Mycoplasma capricolum subsp. capricolum is a well-known pathogen of small ruminants. A recent human case of septicemia involving this agent raised the question of its potential pathogenicity to humans. This spherical organism is distinguished from other bacteria by its small size (a characteristic of the genus Mycoplasma) and requirement of cholesterol for growth. However, its DNA structure suggests that Mycoplasma capricolum is derivative of Gram-positive bacteria
Mycoplasma capricolum subsp. capricolum is a known etiologic agent of contagious agalactia in small ruminants, a disease associated with chronic inflammation, arthritis, and mastitis (1). Even though M. capricolum subsp. capricolum is considered one of the least pathogenic members of the Mycoplasma mycoides cluster, disease outbreaks caused by this agent can have a significant impact on goat farming industries due to loss of milk production and increased mortality
The ribosomal protein gene cluster of Mycoplasma capricolum :
The DNA sequence of the part of the Mycoplasma capricolum genome that contains the genes for 20 ribosomal proteins and two other proteins has been determined. The organization of the gene cluster is essentially the same as that in the S10 and spc operons of Escherichia coli. The deduced amino acid sequence of each protein is also well conserved in the two bacteria. The G+C content of the M. capricolum genes is 29%, which is much lower than that of E. coli (51%). The codon usage pattern of M. capricolum is different from that of E. coli and extremely biased to use of A and U(T): about 91% of codons have A or U in the third position. UGA, which is a stop codon in the “universal” code, is used more abundantly than UGG to dictate tryptophan.
A change in the genetic code inMycoplasma capricolum :
Mycoplasma capricolum was previously found to use UGA instead of UGG as its codon for tryptophan and to contain 75%A+ T in its DNA. The codon change could have been due to mutational pressure to replace C+G by A+T, resulting in the replacement of UGA stop codons by UAA, change of the anticodon in tryptophan tRNA from CCA to UCA, and replacement of UGG tryptophan codons by UGA. None of these changes should have been deleterious.
protein composition:
The whole cell proteins and the ribosomal proteins of Mycoplasma capricolum ATCC 27343 have been analyzed by two-dimensional polyacrylamide gel electrophoresis. The M. capricolum cell is relatively rich in basic proteins. The number of total protein spots detected was approximately 355, which is less than one-third of that of Escherichia coli or Bacillus subtilis. In contrast, the number (30 and 20 protein species have been found to be present in the 50S and 30S ribosomal subunits, respectively) and the size of the ribosomal proteins in the M. capricolum do not seem to be significantly different from those of typical eubacteria.
Codon Bias :
CGG is an arginine codon in the universal genetic code. We previously reported that in Mycoplasma capricolum, a relative of Gram-positive eubacteria, codon CGG did not appear in coding frames, including termination sites, and tRNA(ArgCCG) pairing with codon CGG, was not detected. These facts suggest that CGG is a nonsense (unassigned and untranslatable) codon--i.e., not assigned to arginine or to any other amino acid. We have investigated whether CGG is really an unassigned codon by using a cell-free translation system prepared from M. capricolum. Translation of synthetic mRNA containing in-frame CGG codons does not result in "read-through" to codons beyond the CGG codons--i.e., translation ceases just before CGG. Sucrose-gradient centrifugation profiles of the reaction mixture have shown that the bulk of peptide that has been synthesized is attached to 70S ribosomes and is released upon further incubation with puromycin. The result suggests that the peptide is in the P site of ribosome in the form of peptidyl-tRNA, leaving the A site empty. When in-frame CGG codons are replaced by UAA codons in mRNA, no read-through occurs beyond UAA, just as in the case of CGG. However, the synthesized peptide is released from 70S ribosomes, presumably by release factor 1. These data suggest strongly that CGG is an unassigned codon and differs from UAA in that CGG is not used for termination.
Nucleotide sequence :
The nucleotide sequences of the rrnB 16S ribosomal RNA gene and its 5'-and 3'-flanking regions from Mycoplasma capricolum have been determined. The coding sequence is 1521 base pairs long, being 21 base pairs shorter than that of the Escherichia coli 16S rRNA gene. The 16S rRNA sequence of M. capricolum reveals 74% and 76% identify with that of E. coli and Anacystis nidulans, respectively. The secondary structure model constructed from the M. capricolum 16S rRNA gene sequence resembles that proposed for E. coli 16S rRNA. A large stem structure can be constructed between the 5'- and 3'-flanking sequences of the 16S rRNA gene. The flanking regions are extremely rich in AT.
Mycoplasma capricolum genome ;
parasitic eubacterium Mycoplasma capricolum sequenced by genomic walking techniques. The 287 putative proteins detected to date represent about half of the estimated total number of 500 predicted for this organism. A large fraction of these (75%) can be assigned a likely function as a result of similarity searches. Several important features of the functional organization of this small genome are already apparent. Among these are (i) the expected relatively large number of enzymes involved in metabolic transport and activation, for efficient use of host cell nutrients; (ii) the presence of anabolic enzymes; (iii) the unexpected diversity of enzymes involved in DNA replication and repair: and (iv) a sizeable number of orthologues (82 so far) in Escherichia coli. This survey is beginning to provide a detailed view of how M. capricolum manages to maintain essential cellular processes with a genome much smaller than that of its bacterial relatives.