skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.


Title: A-to-I mRNA editing controls spore death induced by a fungal meiotic drive gene in homologous and heterologous expression systems
Abstract Spore killers are meiotic drive elements that can block the development of sexual spores in fungi. In the maize ear rot and mycotoxin-producing fungus Fusarium verticillioides, a spore killer called SkK has been mapped to a 102-kb interval of chromosome V. Here, we show that a gene within this interval, SKC1, is required for SkK-mediated spore killing and meiotic drive. We also demonstrate that SKC1 is associated with at least 4 transcripts, 2 sense (sense-SKC1a and sense-SKC1b) and 2 antisense (antisense-SKC1a and antisense-SKC1b). Both antisense SKC1 transcripts lack obvious protein-coding sequences and thus appear to be noncoding RNAs. In contrast, sense-SKC1a is a protein-coding transcript that undergoes A-to-I editing to sense-SKC1b in sexual tissue. Translation of sense-SKC1a produces a 70-amino-acid protein (Skc1a), whereas the translation of sense-SKC1b produces an 84-amino-acid protein (Skc1b). Heterologous expression analysis of SKC1 transcripts shows that sense-SKC1a also undergoes A-to-I editing to sense-SKC1b during the Neurospora crassa sexual cycle. Site-directed mutagenesis studies indicate that Skc1b is responsible for spore killing in Fusarium verticillioides and that it induces most meiotic cells to die in Neurospora crassa. Finally, we report that SKC1 homologs are present in over 20 Fusarium species. Overall, our results demonstrate that fungal meiotic drive elements like SKC1 can influence the outcome of meiosis by hijacking a cell’s A-to-I editing machinery and that the involvement of A-to-I editing in a fungal meiotic drive system does not preclude its horizontal transfer to a distantly related species.  more » « less
Award ID(s):
2005295
PAR ID:
10342586
Author(s) / Creator(s):
; ; ; ; ;
Editor(s):
Freitag, M
Date Published:
Journal Name:
Genetics
Volume:
221
Issue:
1
ISSN:
1943-2631
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. Meiotic drive elements, sometimes called selfish genes, are genetic elements that are passed on to their offspring more favorably than other genes. Meiotic drive elements have been observed in many organisms, including fungi of the Neurospora genus. Three different meiotic drive elements, called Spore killers, have been identified in Neurospora fungi. One of these Spore killers is called Spore killer 3 (Sk-3), and the molecular mechanism by which Sk-3 acts as a meiotic drive element is poorly understood. Previous work has identified a genetic locus within Sk-3 that may control spore killing. In this thesis, I investigate an interval of DNA within this locus called v375. Through gene deletion and spore killing assays, I show that deletion of v375 disrupts Sk-3-based spore killing in N. crassa. Possible explanations for why interval v375 is required for spore killing by Sk-3 are discussed. 
    more » « less
  2. Neurospora crassa is a well-known model organism for studying eukaryotic genetics, particularly non-Mendelian inheritance mechanisms such as meiotic drive. In N. crassa, meiotic drive can be observed in fungal spore killing, where Spore killer-3 (Sk-3) is a selfish genetic element transmitted to offspring through spore killing. Sk-3 is thought to contain two principal components: a killer (poison) gene and a resistance (antidote) gene. While the resistance gene (rsk) has been identified, the killer gene remains unknown. Building on previous research that identified a 1.3 kb DNA interval (i350) essential for Sk-3-based spore killing, I analyzed two subintervals, i382 and i400, to narrow down the functional components of the Sk-3 locus. Deletion of i382 does not disrupt spore killing and deletion of interval i400 partially disrupts spore killing but does not eliminate it. Future work should retest the i400 strains in spore killing assays, to determine if the partial spore killing phenotype can be detected in all crosses when larger numbers of rosettes are examined. The findings presented here help narrow down the search for the unknown poison gene involved in spore killing, which is a critical step towards understanding processes that allow for the evolution of Sk-3-type selfish genetic elements. 
    more » « less
  3. Meiotic drive is a non-Mendelian inheritance phenomenon where selfish genetic elements change gene transmission in their own favor. This phenomenon occurs in the fungus Neurospora crassa during spore killing. When a strain carrying a spore killer genetic element is crossed with a non-spore killing wild type strain, the cross will produce half viable and half inviable offspring. The N. crassa Sk-3 spore killer is found on Chromosome III. Sk-3 is one of the most studied meiotic drive elements in Neurospora fungi and it is thought to require a killer gene and a resistance gene for spore killing. While the killer gene has not been identified, recent work has isolated a mutation (rfk-2UV) that disrupts spore killing. Although this mutation has been mapped to Chromosome III, its exact location is not known. In this work, I investigate the role of one DNA interval in Sk-3-based spore killing. This DNA interval, referred to as v373, is thought to reside within or near rfk-2UV. My results will contribute to future efforts to identify the Sk-3 killer gene. 
    more » « less
  4. ABSTRACT NeurosporaSk-2is a complex meiotic drive element that is transmitted to offspring through sexual reproduction in a biased manner.Sk-2’s biased transmission mechanism involves spore killing, and recent evidence has demonstrated that spore killing is triggered by a gene calledrfk-1. However, a second gene,rsk, is also critically important for meiotic drive by spore killing because it allows offspring with anSk-2genotype to survive the toxic effects ofrfk-1. Here, we present evidence demonstrating thatrfk-1encodes two protein variants: a 102 amino acid RFK-1Aand a 130 amino acid RFK-1B, but only RFK-1Bis toxic. We also show that expression of RFK-1Brequires an early stop codon inrfk-1mRNA to undergo adenosine-to-inosine (A-to-I) mRNA editing. Finally, we demonstrate that RFK-1Bis toxic when expressed within vegetative tissue of Spore killer sensitive (SkS) strains, and that this vegetative toxicity can be overcome by co-expressingSk-2’s version of RSK. Overall, our results demonstrate thatSk-2uses RNA editing to control when its spore killer is produced, and that the primary killing and resistance functions ofSk-2can be conferred upon anSkSstrain by the transfer of only two genes. 
    more » « less
  5. Meiotic drivers are selfish genetic elements that skew transmission in their favor. In the filamentous fungus N. crassa, one such meiotic driver is Spore killer-3 (Sk-3). In a cross between Sk-3 and a spore killer-sensitive mating partner (Sk-S), only half of the ascospores (sexual spores) survive. Nearly all of the survivors inherit the genes for spore killing. Previous studies have established that a gene called rfk-2 (required for spore killing) is essential for the spore killing activity of Sk-3. The rfk-2 gene has been mapped to Chromosome III, but its exact location is unknown. The goal of this study is to help identify the exact location of rfk-2. Towards this goal, I investigated a DNA interval called v378. Preliminary findings suggested that this interval may be important for spore killing. To determine if v378 is required for spore killing, I constructed and used a transformation vector to replace v378 with a hygromycin resistance gene (hph+) in an N. crassa Sk-3 strain. Strains deleted of v378 were then crossed with two spore killing-sensitive tester strains. The spore sacs containing ascospores from these crosses were imaged to analyze the effects of replacement of v378 on Sk-3-based spore killing. My findings demonstrate that v378 is required for spore killing. The potential implications of my findings with respect to our understanding of meiotic drive elements, and their potential applications, is discussed. 
    more » « less