Unknown,Transcriptomics,Genomics,Proteomics

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Differential gene expression in a Cryptococcus neoformans ccr4Delta mutant during host-temperature adaptation

ABSTRACT: Analysis of ccr4 Delta vs. wild type shifted from 30oC to 37oC for 10 minutes To determine the effect of ccr4 deletion on temperature stress-responsive changes in gene expression in C. neoformans Replicate cultures of the wild type (Control) and ccr4Delta mutant were grown to mid log phase at 30oC, harvested and resuspended in 37oC pre-warmed medium for 10 minutes. Pooled biological replicate cultures were hybridized in duplicate (2 slides, 2 arrays/slide)

ORGANISM(S): Cryptococcus neoformans

SUBMITTER: John Panepinto

PROVIDER: E-GEOD-28592 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The transmission of the malaria parasite between mosquitoes and mammals requires translational repression to ensure that only the proper proteins are expressed at the right time, while still allowing the parasite to prepare the mRNAs it will need for the next developmental stage. With relatively few known specific transcription factors (ApiAP2 family) that may specifically initiate gene transcription, Plasmodium parasites also regulate the stability and turnover of transcripts to provide more comprehensive gene regulation. The CAF1/CCR4/NOT complex has been shown in model organisms to be important for not only mRNA degradation, but also translational control through its deadenylases CAF1 and CCR4. However, few proteins that impose translational repression in Plasmodium sexual stages are known, and those that are characterized primarily affect female gametocytes. Therefore, we have characterized two deadenylases and have uncovered their roles in transmission. We have identified and characterized CCR4-1, which we show plays a role in activating male gametocytes, stabilizing transcripts in gametocytes, and regulating host-to-vector transmission. We find that when ccr4-1 is genetically deleted, there is a loss in the coordination of male gametocyte activation and a reduction in the ability of the parasite to productively infect the mosquito, which is independent of its effect upon male activation. Comparative RNA-seq shows that the deletion of ccr4-1 affects many transcripts that are translationally repressed in females, are related to male gamete function, and/or are important for early mosquito stage development. In contrast, we found that genetic deletion of the major deadenylase Caf1 is lethal. However, we observed that expression of only the N-terminal Caf1 domain is permissive, yet prevents proper complex assembly and phenocopies the ccr4-1 deletion. We therefore conclude that the general and transmission-specialized deadenylases of the CAF1/CCR4/NOT complex play critical and intertwined roles in parasite growth and transmission.

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Differential gene expression in a Cryptococcus neoformans ccr4Delta mutant during host-temperature adaptation

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