Involvement of cDNA in Homologous Recombination between Ty Elements in <i>Saccharomyces cerevisiae</i>

Melamed, C; Nevo, Yoram; Kupiec, Martin

doi:10.1128/mcb.12.4.1613-1620.1992

Cited by 12 publications

(8 citation statements)

References 33 publications

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“…S4D). We conclude that the Gag chimeras support de novo retrotransposition and cDNA recombination remains a minor pathway (64,65).…”

Section: Ty1 Mobility Of Gag Chimeras Containing Foreign Prldsmentioning

confidence: 78%

An interchangeable prion-like domain is required for Ty1 retrotransposition

Beckwith¹,

Nomberg²,

Newman³

et al. 2023

Preprint

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Retrotransposons and retroviruses shape genome evolution and can negatively impact genome function.Saccharomyces cerevisiaeand its close relatives harbor several families of LTR-retrotransposons, the most abundant being Ty1 in several laboratory strains. The cytosolic foci that nucleate Ty1 virus-like particle (VLP) assembly are not well-understood. These foci, termed retrosomes or T-bodies, contain Ty1 Gag and likely Gag-Pol and the Ty1 mRNA destined for reverse transcription. Here, we report a novel intrinsically disordered N-terminal prion-like domain (PrLD) within Gag that is required for transposition. This domain contains amino-acid composition similar to known yeast prions and is sufficient to nucleate prionogenesis in an established cell-based prion reporter system. Deleting the Ty1 PrLD results in dramatic VLP assembly and retrotransposition defects but does not affect Gag protein level. Ty1 Gag chimeras in which the PrLD is replaced with other sequences, including yeast and mammalian prionogenic domains, display a range of retrotransposition phenotypes from wildtype to null. We examine these chimeras throughout the Ty1 replication cycle and find that some support retrosome formation, VLP assembly, and retrotransposition, including the yeast Sup35 prion and the mouse PrP prion. Our interchangeable Ty1 system provides a useful, genetically tractablein vivoplatform for studying PrLDs, complete with a suite of robust and sensitive assays, and host modulators developed to study Ty1 retromobility. Our work invites study into the prevalence of PrLDs in additional mobile elements.

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“…S4D). We conclude that the Gag chimeras support de novo retrotransposition and cDNA recombination remains a minor pathway (64,65).…”

Section: Ty1 Mobility Of Gag Chimeras Containing Foreign Prldsmentioning

confidence: 78%

An interchangeable prion-like domain is required for Ty1 retrotransposition

Beckwith¹,

Nomberg²,

Newman³

et al. 2023

Preprint

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“…Experiments in our laboratory have shown that gene conversion between homologous sequences located at different positions in the genome (ectopic gene conversion) is induced by UV light in a similar way to allelic gene conversion (17a; also see below). Since recombination between Ty elements seems to follow different rules from those seen for other sequences (18,24), we decided to check whether Ty-Ty recombination could be induced by UV irradiation.…”

Section: Resultsmentioning

confidence: 99%

“…Strain AP5, carrying an ectopic copy of the lys2::URA3 allele, was constructed by transforming AP1 with pAP5 to His' Ura+. Strains MK95 and MK104 were 4441 created by transforming MK89 (A4Ta ura3-Nco-his3-11, 15 trpl-Xba-canl-101) to Ura+ Lys-with plasmids pM87 and pM115, respectively (24). These two strains differ at the 5' long terminal repeat (LTR) of the TylUra located at the LYS2 locus.…”

Section: Methodsmentioning

confidence: 99%

Ectopic Recombination between Ty Elements in Saccharomyces cerevisiae Is Not Induced by DNA Damage

Parket

Kupiec

1992

Molecular and Cellular Biology

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Mitotic recombination is increased when cells are treated with a variety of physical and chemical agents that cause damage to their DNA. We show here, using Saccharomyces cerevisiae strains that carry marked Ty elements, that recombination between members of this family of retrotransposons is not increased by UV irradiation or by treatment with the radiomimetic drug methyl methanesulfonate. Both ectopic recombination and mutation events were elevated by these agents for non-Ty sequences in the same strain. We discuss possible mechanisms that can prevent the induction of recombination between Ty elements.

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“…While the genetic effects of integration have been well documented, it has become increasingly clear that recombination of retrotransposon cDNA into the genome can also shape the genetic material. Studies with Tyl elements have shown that Tyl cDNA can recombine with preexisting insertions to generate simple replacements or to form tandem elements (WEINSTOCK et MELAMED et al 1992;SHARON et al 1994). Cellular mRNA may also occasionally get reverse transcribed by retroelements.…”

Section: Discussionmentioning

confidence: 99%

“…Genetically marked Tyl cDNA that results from reverse transcription typically enters the yeast genome by the integration pathway. This cDNA can also recombine with a chromosomal Tyl element such that the chromosomal element acquires the cDNA marker gene (MELAMED et al 1992). In addition, Tyl cDNA recombination can yield tandem elements, and they are frequently observed when the integration pathway is blocked (SHARON et al 1994).…”

mentioning

confidence: 99%

High Frequency cDNA Recombination of the Saccharomyces Retrotransposon Ty5: The LTR Mediates Formation of Tandem Elements

Ning

Voytas

1997

Genetics

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Retroelement cDNA can integrate into the genome using the element-encoded integrase, or it can recombine with preexisting elements using the recombination system of the host. Recombination is a particularly important pathway for the yeast retrotransposon Ty5 and accounts for ∼30% of the putative transposition events when a homologous substrate is carried on a plasmid and ∼7% when the substrate is located at the chromosomal URA3 locus. Characterization of recombinants revealed that they are either simple replacements of the marker gene or tandem elements. Using an assay system in which the donor element and recombination substrates are separated, we found that the long terminal repeats (LTRs) are critical for tandem element formation. LTR-containing substrates generate tandem elements at frequencies more than 10-fold higher than similarly sized internal Ty5 sequences. Internal sequences, however, facilitate tandem element formation when associated with an LTR, and there is a linear relationship between frequencies of tandem element formation and the length of LTR-containing substrates. We propose that recombination is initiated between the LTRs of the cDNA and substrate and that internal sequences promote tandem element formation by facilitating sequence alignment. Because of its location in subtelomeric regions, recombinational amplification of Ty5 may contribute to the organization of chromosome ends.

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Involvement of cDNA in Homologous Recombination between Ty Elements in Saccharomyces cerevisiae

Cited by 12 publications

References 33 publications

An interchangeable prion-like domain is required for Ty1 retrotransposition

An interchangeable prion-like domain is required for Ty1 retrotransposition

Ectopic Recombination between Ty Elements in Saccharomyces cerevisiae Is Not Induced by DNA Damage

High Frequency cDNA Recombination of the Saccharomyces Retrotransposon Ty5: The LTR Mediates Formation of Tandem Elements

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