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Volume 156, Issue 2

[poly(A) polymerase 1] homozygosity and hyperadenylation are major determinants of increased mRNA stability of in azole-resistant clinical isolates of Free

Abstract

Using genetically matched azole-susceptible (AS) and azole-resistant (AR) clinical isolates of , we recently demonstrated that overexpression in AR isolates is due to its enhanced transcriptional activation and mRNA stability. This study examines the molecular mechanisms underlying enhanced mRNA stability in AR isolates. Mapping of the 3′ untranslated region (3′ UTR) of revealed that it was rich in adenylate/uridylate (AU) elements, possessed heterogeneous polyadenylation sites, and had putative consensus sequences for RNA-binding proteins. Swapping of heterologous and chimeric – 3′ UTR transcriptional reporter fusion constructs did not alter the reporter activity in AS and AR isolates, indicating that -acting sequences within the 3′ UTR itself are not sufficient to confer the observed differential mRNA decay. Interestingly, the poly(A) tail of the mRNA of AR isolates was ∼35–50 % hyperadenylated as compared with AS isolates. poly(A) polymerase (), responsible for mRNA adenylation, resides on chromosome 5 in close proximity to the mating type-like () locus. Two different alleles, , were recovered from AS (), while a single type of allele () was recovered from AR isolates (). Among the heterozygous deletions of (Δ) and (Δ), only the former led to relatively enhanced drug resistance, to polyadenylation and to transcript stability of in the AS isolate. This suggests a dominant negative role of in transcript polyadenylation and stability. Taken together, our study provides the first evidence, to our knowledge, that loss of heterozygosity at the locus is linked to hyperadenylation and subsequent increased stability of transcripts, thus contributing to enhanced drug resistance.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): 3′ UTR, 3′ untranslated region , AR, azole-resistant , AS, azole-susceptible , LOH, loss of heterozygosity , MDR, multidrug resistance , NouR, nourseothricin-resistant , PAT, polyadenylation test and RACE, rapid amplification of cDNA ends
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2010-02-01
2024-04-18
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PAP1 [poly(A) polymerase 1] homozygosity and hyperadenylation are major determinants of increased mRNA stability of CDR1 in azole-resistant clinical isolates of Candida albicans
Microbiology 156, 313 (2010); https://doi.org/10.1099/mic.0.035154-0
/content/journal/micro/10.1099/mic.0.035154-0
/content/journal/micro/10.1099/mic.0.035154-0
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