The
wild-type
DNA
sequence 5'- CTA
CAG ATT - 3'
3'- GAT GTC TAA
-
5'
produces the mRNA
5'- CUA CAG AUU
-
3'
which codes for the
polypeptide
- leu-gln-iso -
[etc]
A 1st position mutation (CT) in the second triplet gives a 'mutant' DNA sequence
sense
strand
5'- CTA AG
ATT - 3'
3'- GAT TC
TAA - 5'
produces the mRNA
5'- CUA AG
AUU - 3'
which codes for the
polypeptide
- leu- *
because UAG is a 'stop' codon (the so-called amber stop). Chain growth in the polypeptide terminates prematurely.
The portion of the transfer RNA gene (tDNA) that codes for the anticodon loop of tRNAtyr(UAC) , which recognizes the mRNA codon 'UAC' and inserts 'tyr', is
tDNA
gene
3'- ATG -5'
5'- TAC -3'
which produces the tRNA
3'-
AUG -5' anticodon
loop
which reads the mRNA
5'- UAC -3'
as
- tyr -
A 3rd-position mutation (CG) on the 5'3' strand in this region produces a 'mutant' tDNA gene
tDNA
gene
3'- AT
-5'
5'- TA
-3'
which produces the tRNA
3'-
AU
-5' anticodon loop
which reads the mRNA
5'- UAG -3'
as
- tyr -
Then, the mutated tDNA gene produces a tRNA molecule that will read the the mutated gln codon UAG in part 1 as a tyr codon, instead of as a 'stop' codon.
The polypeptide
will have an amino acid
substitution (glntyr);
translation
of
the rest of the mRNA will proceed normally, in the
correct reading frame. The mutation in the tDNA gene is
said to
'suppress' the phenotypic effect of the first
mutation in
a protein-coding gene, and the supressor
tRNA may allow enough of the mRNA to be
translated
to permit a 'standard' phenotype. Note that other 'stop'
codons
will
be read incorrectly as 'tyr': however, most will be read
by
'standard'
tRNAs and the protein
products will be terminated normally.