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The remarkable capacity of neurons to reorganize their structure, function, and connections in an activity-dependent manner is supported by an extensive network of molecules and effectors. Among those, Activity-regulated cytoskeleton-associated protein (Arc, also known as Arg3.1) is considered one of the central, most versatile, players. Arc is an immediate early gene product that support neuroplastic changes important for cognitive function and memory formation, while also oligomerizing to form capsids that can be released and can move independent of the synapse. Although a lot has been uncovered about the contributions of Arc to neuron biology and behavior, very little is known about how different functions of Arc are regulated both temporally and spatially in neurons. To answer to this question, the regulation of Arc expression must be monitored at various levels. This thesis analyzes how Arc can be transcriptionally and post-translationally regulated and how this confers neurobiological specificity. Being an immediate early gene, transcription of Arc needs to act rapidly in response to a neuronal stimulus, such as BDNF. The first aim of this thesis provides a model to describe how the Arc promoter can overcome topological constraints to facilitate rapid and persistent transcription, up to 6 hours post-BDNF exposure. Using chromatin analysis and biochemical techniques we have mapped regions of the Arc promoter that undergo double stranded DNA breaks to facilitate rapid transcription, and that BDNF-induced Arc non-coding RNA elements interact with DNA repair proteins such as Brd4 to effectively end transcription. The second aim of this thesis aims to determine how phosphorylation can spatiotemporally dictate the function of Arc. Mass spectrometry and sequence prediction strategies were used to map novel Arc phosphorylation sites. This approach led us to recognize S67 and T278 as residues that can be modified by TNIK, which is a kinase abundantly expressed in neurons and been implicated as a risk factor for psychiatric disorders. Characterization of TNIK-dependant phosphorylation at these two sites can strongly influence Arc's subcellular distribution and self-assembly as capsids. The multifaceted nature of Arc’s neurobiology ranges from being a key component of long-term memory consolidation to participating in viral capsid intracellular communication. This thesis provides insight into Arc’s moonlighting capabilities both at the transcription and post-translational levels.

Estrogens have been implicated extensively in learning, memory, dendritic spinogenesis, and synaptic plasticity. The molecular pathways underlying estrogens’ rapid, nongenomic actions are not well understood, however protein translation of dendritically-localized mRNA transcripts may be a possible mechanism of action. The Activity-regulated cytoskeleton-association protein (Arc) has emerged as a strong candidate for the mediation of these effects. Here, we present findings from mouse in vivo and primary cortical cell culture models demonstrating Arc’s dynamic regulation following stimulation by 17beta-estradiol (E2) within 15- to 40-minutes of treatment. Further, we show that E2 induces a differential localization of dendritic Arc dependent on the length of time that follows E2 treatment. In sum, these data suggest a novel role for Arc acting within a mechanism of E2-facilitated neuronal actions that is dose-, time-, and dendritic region-dependent, advocating for a multi-mechanism approach to studying Arc’s involvement in estrogenic short-term memory facilitation within the mouse brain.