RNA-Seq analysis of wild-type and ttpA-C240ins mutant D. discoideum amebae during short-term nutrient deprivation
In many eukaryotes, mRNAs containing specific AU-rich motifs are regulated by proteins of the tristetraprolin (TTP) family, which bind to these motifs through a tandem zinc finger (TZF) domain. This binding leads to promotion of subsequent deadenylation and decay, partly through a conserved carboxyl-terminal CNOT1 binding domain. We explored the physiological role of the single TTP family member (TtpA) expressed in Dictyostelium discoideum. This study shows the effect of an insertional mutation in the TZF domain (ttpA-C240ins), which disrupted the TZF RNA binding domain, evaluated in amebae in axenic conditions in growth medium during surface culture, and during short term (2 and 6 hours) nutrient deprivation. RNA-Seq analysis was performed on WT and mutant cells in parallel during this short time course. Overall design: Dictyostelium discoideum amebae (strain AX3) were grown in normal growth medium under axenic conditions on plastic surfaces until they were approximately 80% confluent. Three cultures of wild-type cells and 8-9 cultures of mutant cells were then frozen and used for RNA extraction and RNA-Seq analysis. Other cultures of the same type were changed to nutrient-deficient medium, and harvested after 2 and 6 hours.
External Link: /pubmed:34718768