A small targeting domain in Ty1 integrase is sufficient to direct retrotransposon integration upstream of tRNA genes.

The EMBO Journal
Amna Asif-LaidinPascale Lesage

Abstract

Integration of transposable elements into the genome is mutagenic. Mechanisms targeting integrations into relatively safe locations, hence minimizing deleterious consequences for cell fitness, have emerged during evolution. In budding yeast, integration of the Ty1 LTR retrotransposon upstream of RNA polymerase III (Pol III)-transcribed genes requires interaction between Ty1 integrase (IN1) and AC40, a subunit common to Pol I and Pol III. Here, we identify the Ty1 targeting domain of IN1 that ensures (i) IN1 binding to Pol I and Pol III through AC40, (ii) IN1 genome-wide recruitment to Pol I- and Pol III-transcribed genes, and (iii) Ty1 integration only at Pol III-transcribed genes, while IN1 recruitment by AC40 is insufficient to target Ty1 integration into Pol I-transcribed genes. Swapping the targeting domains between Ty5 and Ty1 integrases causes Ty5 integration at Pol III-transcribed genes, indicating that the targeting domain of IN1 alone confers Ty1 integration site specificity.

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Citations

Feb 24, 2021·Nucleic Acids Research·Angelika AndrzejewskaKatarzyna Pachulska-Wieczorek

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Methods Mentioned

BETA
fluorescence microscopy
immunoprecipitation
ChIP
PCR
environmental stress

Software Mentioned

Deeptools
MarkDuplicates
BWA short
FasqtQC
Genome
R
Bowtie2
Andor IQ2
Bigwig

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