Host factors that promote retrotransposon integration are similar in distantly related eukaryotes

Autor: Maya Sangesland, Sudhir K. Rai, Yujin Cui, Caroline Esnault, Henry L. Levin, Atreyi Ghatak Chatterjee, Michael Lee
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
Cancer Research
DNA Repair
Gene Expression
Retrotransposon
Yeast and Fungal Models
Biochemistry
Schizosaccharomyces Pombe
Retrovirus
Gene Expression Regulation
Fungal
Promoter Regions
Genetic
Homologous Recombination
Genetics (clinical)
Genetics
Recombination
Genetic
biology
Chromosome Biology
Eukaryota
RNA-Directed DNA Polymerase
Long terminal repeat
Recombinant Proteins
Chromatin
Nucleic acids
Recombination-Based Assay
Experimental Organism Systems
Epigenetics
Schizosaccharomyces
Research Article
Integration Host Factors
Saccharomyces cerevisiae Proteins
Retroelements
lcsh:QH426-470
DNA recombination
Ubiquitin-Protein Ligases
Saccharomyces cerevisiae
Library Screening
Research and Analysis Methods
03 medical and health sciences
Saccharomyces
Model Organisms
Complementary DNA
DNA-binding proteins
Molecular Biology Techniques
Gene
Molecular Biology
Ecology
Evolution
Behavior and Systematics
Molecular Biology Assays and Analysis Techniques
030102 biochemistry & molecular biology
Integrases
Biology and life sciences
Terminal Repeat Sequences
Organisms
Fungi
Proteins
Promoter
DNA
Cell Biology
biology.organism_classification
Yeast
lcsh:Genetics
030104 developmental biology
Retroviridae
Zdroj: PLoS Genetics, Vol 13, Iss 12, p e1006775 (2017)
PLoS Genetics
ISSN: 1553-7404
1553-7390
Popis: Retroviruses and Long Terminal Repeat (LTR)-retrotransposons have distinct patterns of integration sites. The oncogenic potential of retrovirus-based vectors used in gene therapy is dependent on the selection of integration sites associated with promoters. The LTR-retrotransposon Tf1 of Schizosaccharomyces pombe is studied as a model for oncogenic retroviruses because it integrates into the promoters of stress response genes. Although integrases (INs) encoded by retroviruses and LTR-retrotransposons are responsible for catalyzing the insertion of cDNA into the host genome, it is thought that distinct host factors are required for the efficiency and specificity of integration. We tested this hypothesis with a genome-wide screen of host factors that promote Tf1 integration. By combining an assay for transposition with a genetic assay that measures cDNA recombination we could identify factors that contribute differentially to integration. We utilized this assay to test a collection of 3,004 S. pombe strains with single gene deletions. Using these screens and immunoblot measures of Tf1 proteins, we identified a total of 61 genes that promote integration. The candidate integration factors participate in a range of processes including nuclear transport, transcription, mRNA processing, vesicle transport, chromatin structure and DNA repair. Two candidates, Rhp18 and the NineTeen complex were tested in two-hybrid assays and were found to interact with Tf1 IN. Surprisingly, a number of pathways we identified were found previously to promote integration of the LTR-retrotransposons Ty1 and Ty3 in Saccharomyces cerevisiae, indicating the contribution of host factors to integration are common in distantly related organisms. The DNA repair factors are of particular interest because they may identify the pathways that repair the single stranded gaps flanking the sites of strand transfer following integration of LTR retroelements.
Author summary Retroviruses and retrotransposons are genetic elements that propagate by integrating into chromosomes of eukaryotic cells. Genetic disorders are being treated with retrovirus-based vectors that integrate corrective genes into the chromosomes of patients. Unfortunately, the vectors can alter expression of adjacent genes and depending on the position of integration, cancer genes can be induced. It is therefore essential that we understand how integration sites are selected. Interestingly, different retroviruses and retrotransposons have different profiles of integration sites. While specific proteins have been identified that select target sites, it’s not known what other cellular factors promote integration. In this paper, we report a comprehensive screen of host factors that promote LTR-retrotransposon integration in the widely-studied yeast, Schizosaccharomyces pombe. Unexpectedly, we found a wide range of pathways and host factors participate in integration. And importantly, we found the cellular processes that promote integration relative to recombination in S. pombe are the same that drive integration of LTR-retrotransposons in the distantly related yeast Saccharomyces cerevisiae. This suggests a specific set of cellular pathways are responsible for integration in a wide range of eukaryotic hosts.
Databáze: OpenAIRE
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