PT  - JOURNAL ARTICLE
AU  - Tho Huu Nguyen
AU  - Tae Hee Han
AU  - Stuart Newfeld
AU  - Mihaela Serpe
TI  - Selective disruption of synaptic BMP signaling by a Smad mutation adjacent to the highly conserved H2 helix
AID  - 10.1101/811109
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 811109
4099  - http://biorxiv.org/content/early/2019/10/21/811109.short
4100  - http://biorxiv.org/content/early/2019/10/21/811109.full
AB  - Bone morphogenetic proteins (BMPs) shape normal development and function via canonical and non-canonical signaling pathways. When activating the canonical pathway, BMPs initiate signaling by binding to transmembrane receptors that phosphorylate pathway effectors, the Smad proteins, inducing their translocation into the nucleus and thus regulation of target genes. Phosphorylated Smads also accumulate at cellular junctions, but this non-canonical signaling modality remains less defined. We have recently reported that phosphorylated Smad (pMad in Drosophila) accumulates at synaptic junctions in complexes with genetically distinct composition and regulation. Here we examined a wide collection of Drosophila Mad alleles and searched for molecular features relevant to pMad accumulation at synaptic junctions. We found that strong Mad alleles generally disrupt both synaptic and nuclear pMad accumulation, whereas moderate Mad alleles have a wider range of phenotypes and could selectively impact different BMP signaling modalities. Interestingly, synaptic pMad appeared more sensitive to net reduction in Mad levels than nuclear pMad. Importantly, a previously uncharacterized allele, Mad8, showed markedly reduced synaptic pMad levels but only moderately diminished nuclear pMad signals. The postsynaptic composition and electrophysiological properties of Mad8 NMJs were similarly altered. Using biochemical approaches, we examined how single point mutations such as S359L, present in Mad8, could influence the Mad-receptor interface and we identified a key molecular determinant, the H2 helix. Our study highlights the biological relevance of the Smad-dependent, non-canonical BMP signaling and uncovers a highly conserved structural feature of Smads, critical for normal development and function.