title: Accidental amplification and inactivation of a methyltransferase gene eliminates cytosine methylation in Mycosphaerella graminicola.
abstract: A de novo search for repetitive elements in the genome sequence of the wheat pathogen Mycosphaerella graminicola identified a family of repeats containing a DNA cytosine methyltransferase sequence (MgDNMT). All 23 MgDNMT sequences identified carried signatures of repeat induced point mutation (RIP). All copies were subtelomeric in location except for one on chromosome 6. Synteny with M. fijiensis implied that the nontelomeric copy on chromosome 6 served as a template for subsequent amplifications. Southern analysis revealed that the MgDNMT sequence also was amplified in 15 additional M. graminicola isolates from various geographical regions. However, this amplification event was specific to M. graminicola; a search for MgDNMT homologs identified only a single, unmutated copy in the genomes of 11 other ascomycetes. A genome-wide methylation assay revealed that M. graminicola lacks cytosine methylation, as expected if its MgDNMT gene is inactivated. Methylation was present in several other species tested, including the closest known relatives of M. graminicola, species S1 and S2. Therefore, the observed changes most likely occurred within the past 10,500 years since the divergence between M. graminicola and S1. Our data indicate that the recent amplification of a single-copy MgDNMT gene made it susceptible to RIP, resulting in complete loss of cytosine methylation in M. graminicola.
authors.affiliation: Department of Botany and Plant Pathology, Purdue University, West Lafayette, Indiana 47907, USA.
authors.forename: Braham
authors.initials: B
authors.lastname: Dhillon
authors.forename: Jessica R
authors.initials: JR
authors.lastname: Cavaletto
authors.forename: Karl V
authors.initials: KV
authors.lastname: Wood
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Goodwin
mesh_terms: D001203:Ascomycota / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D003596:Cytosine / Q000378:metabolism*; D019175:DNA Methylation / Q000235:genetics*; D015257:DNA-Cytosine Methylases / Q000235:genetics* / Q000378:metabolism; D005784:Gene Amplification; D020868:Gene Silencing; D016681:Genome, Fungal / Q000235:genetics; D017354:Point Mutation / Q000235:genetics; D018626:Retroelements / Q000235:genetics; D026801:Synteny / Q000235:genetics; D016615:Telomere / Q000235:genetics; D020079:Terminal Repeat Sequences / Q000235:genetics
chemical_list: D018626:Retroelements; D003596:Cytosine; D015257:DNA-Cytosine Methylases