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Translation initiation of IS50R read-through transcripts.

V P Schulz¹, W S Reznikoff

¹Biochemistry Department, University of Wisconsin, Madison 53706.

Journal of Molecular Biology

|September 5, 1991

Summary

Insertion sequences like IS50R transpose infrequently due to low transposase levels. RNA secondary structures in read-through transcripts limit transposase expression, but mutations can overcome this.

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Area of Science:

Molecular Biology
Genetics
Microbiology

Background:

Insertion sequences (IS) and transposons mediate genetic rearrangement.
Transposase expression is a key regulatory factor for transposition frequency.
IS50R and Tn5 exhibit low transposition rates, linked to limited transposase production.

Purpose of the Study:

Investigate the mechanisms limiting transposase expression from read-through transcripts.
Identify genetic mutations that enhance transposase translation from these transcripts.
Determine the role of RNA secondary structures in regulating transposase expression.

Main Methods:

Isolation and characterization of mutations affecting transposase translation.
In vitro analysis of RNA secondary structures.
Assessment of transposase protein synthesis from read-through transcripts.

Main Results:

Read-through transcripts encoding transposase are produced but poorly translated.
Mutations increasing translation often destabilize a predicted RNA secondary structure.
This secondary structure forms in read-through transcripts but not normal ones.
In vitro studies confirm the RNA structure's role and mutation effects.

Conclusions:

RNA secondary structure in the ribosome binding site significantly limits transposase expression from read-through transcripts.
Mutations disrupting this structure enhance transposase translation.
This highlights RNA structure as a critical regulatory element in transposition.

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