Regulated ATF4 persistence in cell cycle control and neurogenesis

A ctivating T ranscription F actor 4 (ATF4) was originally identified as a regulator of viral BLV long terminal repeat protein expression. Since then, its function has expanded to include roles in cellular stress response, embryonic development, and synaptic plasticity. Mice lacking ATF4 generally die at birth and exhibit profound growth retardation with striking developmental defects in the eye and skeletal system, underscoring a crucial role for ATF4 expression during development. While much research has focused on elucidating specific ATF4 target genes in various contexts, very little is known about how ATF4 itself is regulated. Understanding the mechanisms that control ATF4 expression is likely to provide further insight into its function. In this work, I detail the mechanistics surrounding ATF4 degradation and describe a novel mode by which cells can fine tune ATF4-dependent transcription. Steady state ATF4 levels are regulated by a gradient of proline-directed phosphorylation, which in turn converge to regulate phosphorylation of the β-TrCP degron and subsequent ubiquitin-dependent proteolysis. ATF4 levels oscillate during the cell cycle, implying that its expression needs to be kept within a tightly regulated temporal window. ATF4 persistence induces an accumulation of cells in early G1 both in cell lines and neural progenitors in vivo. This cell cycle arrest impairs the process of neurogenesis and neuronal migration. Therefore, proper control of ATF4 dosage is important for bridging consecutive cell cycles, which in turn is required for neural progenitors to efficiently differentiate into neurons. In the second section, I expand on results from the first section to describe a role for cyclin-dependent kinase 5 (CDK5) in regulating ATF4 degradation. CDK5 activity induces ATF4 hyper-phosphorylation, promotes association with β-TrCP, and decreases steady state ATF4 levels. As CDK5 is a constitutively active proline-directed kinase in neurons, this mechanism provides an explanation of how ATF4 levels are kept low in neurons. In addition, increased ATF4 dosage inhibits neurite outgrowth, exemplifying the negative consequences of persistent ATF4 expression in a post-mitotic environment.