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NOVA2-mediated RNA regulation is required for axonal pathfinding during development.

Saito Y, Miranda-Rottmann S, Ruggiu M, Park CY, Fak JJ, Zhong R, Duncan JS, Fabella BA, Junge HJ, Chen Z, Araya R, Fritzsch B, Hudspeth AJ, Darnell RB - Elife (2016)

Bottom Line: The neuron specific RNA-binding proteins NOVA1 and NOVA2 are highly homologous alternative splicing regulators.NOVA proteins regulate at least 700 alternative splicing events in vivo, yet relatively little is known about the biologic consequences of NOVA action and in particular about functional differences between NOVA1 and NOVA2.Thus we have discovered that NOVA2 uniquely regulates alternative splicing of a coordinate set of transcripts encoding key components in cortical, brainstem and spinal axon guidance/outgrowth pathways during neural differentiation, with severe functional consequences in vivo.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Neuro-Oncology, Howard Hughes Medical Institute, The Rockefeller University, New York, United States.

ABSTRACT
The neuron specific RNA-binding proteins NOVA1 and NOVA2 are highly homologous alternative splicing regulators. NOVA proteins regulate at least 700 alternative splicing events in vivo, yet relatively little is known about the biologic consequences of NOVA action and in particular about functional differences between NOVA1 and NOVA2. Transcriptome-wide searches for isoform-specific functions, using NOVA1 and NOVA2 specific HITS-CLIP and RNA-seq data from mouse cortex lacking either NOVA isoform, reveals that NOVA2 uniquely regulates alternative splicing events of a series of axon guidance related genes during cortical development. Corresponding axonal pathfinding defects were specific to NOVA2 deficiency: Nova2-/- but not Nova1-/- mice had agenesis of the corpus callosum, and axonal outgrowth defects specific to ventral motoneuron axons and efferent innervation of the cochlea. Thus we have discovered that NOVA2 uniquely regulates alternative splicing of a coordinate set of transcripts encoding key components in cortical, brainstem and spinal axon guidance/outgrowth pathways during neural differentiation, with severe functional consequences in vivo.

No MeSH data available.


Related in: MedlinePlus

Anti-NOVA2 and anti-NOVA1 antibodies specificity.Coronal brain sections of E16.5 wild-type, Nova1-/-, and Nova2-/-, stained for NOVA1 (green), NOVA2 (red), and DAPI (blue). Scale bar; 500 μm. (B) Specificity of antibodies for immunoprecipitation.DOI:http://dx.doi.org/10.7554/eLife.14371.005
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fig1s2: Anti-NOVA2 and anti-NOVA1 antibodies specificity.Coronal brain sections of E16.5 wild-type, Nova1-/-, and Nova2-/-, stained for NOVA1 (green), NOVA2 (red), and DAPI (blue). Scale bar; 500 μm. (B) Specificity of antibodies for immunoprecipitation.DOI:http://dx.doi.org/10.7554/eLife.14371.005

Mentions: Primary antibodies used for immunohistochemistory and western blot were as follows; goat anti-NOVA2 (C-16) (sc-10546, Santa Cruz), rabbit anti-NOVA1 [EPR13847] (ab183024, abcam), human anti-pan NOVA (anti-Nova paraneoplastic human serum), rabbit anti-PTBP2 (Polydorides et al., 2000), rat anti-L1 (MAB5272, Millipore), goat anti-TAG1/Contactin-2 (AF4439, R&D systems), goat anti-NetrinG1a (AF1166, R&D systems), rabbit anti-NURR1 (M-196) (sc-5568, Santa Cruz), rabbit anti-neurofilament (AB1981, Chemicon), mouse anti-NF200 (N5389, SIGMA), mouse anti-ctbp2 (612044, BD), rabbit anti-myoVI (sc-50461, Santa Cruz), and goat anti-neuropilin-1 (AF566, R&D systems). Anti-NOVA1 and anit-NOVA2 antibody specificity for immunohistochemistory and immunoprecipitation was confirmed (Figure 1—figure supplement 2).


NOVA2-mediated RNA regulation is required for axonal pathfinding during development.

Saito Y, Miranda-Rottmann S, Ruggiu M, Park CY, Fak JJ, Zhong R, Duncan JS, Fabella BA, Junge HJ, Chen Z, Araya R, Fritzsch B, Hudspeth AJ, Darnell RB - Elife (2016)

Anti-NOVA2 and anti-NOVA1 antibodies specificity.Coronal brain sections of E16.5 wild-type, Nova1-/-, and Nova2-/-, stained for NOVA1 (green), NOVA2 (red), and DAPI (blue). Scale bar; 500 μm. (B) Specificity of antibodies for immunoprecipitation.DOI:http://dx.doi.org/10.7554/eLife.14371.005
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4930328&req=5

fig1s2: Anti-NOVA2 and anti-NOVA1 antibodies specificity.Coronal brain sections of E16.5 wild-type, Nova1-/-, and Nova2-/-, stained for NOVA1 (green), NOVA2 (red), and DAPI (blue). Scale bar; 500 μm. (B) Specificity of antibodies for immunoprecipitation.DOI:http://dx.doi.org/10.7554/eLife.14371.005
Mentions: Primary antibodies used for immunohistochemistory and western blot were as follows; goat anti-NOVA2 (C-16) (sc-10546, Santa Cruz), rabbit anti-NOVA1 [EPR13847] (ab183024, abcam), human anti-pan NOVA (anti-Nova paraneoplastic human serum), rabbit anti-PTBP2 (Polydorides et al., 2000), rat anti-L1 (MAB5272, Millipore), goat anti-TAG1/Contactin-2 (AF4439, R&D systems), goat anti-NetrinG1a (AF1166, R&D systems), rabbit anti-NURR1 (M-196) (sc-5568, Santa Cruz), rabbit anti-neurofilament (AB1981, Chemicon), mouse anti-NF200 (N5389, SIGMA), mouse anti-ctbp2 (612044, BD), rabbit anti-myoVI (sc-50461, Santa Cruz), and goat anti-neuropilin-1 (AF566, R&D systems). Anti-NOVA1 and anit-NOVA2 antibody specificity for immunohistochemistory and immunoprecipitation was confirmed (Figure 1—figure supplement 2).

Bottom Line: The neuron specific RNA-binding proteins NOVA1 and NOVA2 are highly homologous alternative splicing regulators.NOVA proteins regulate at least 700 alternative splicing events in vivo, yet relatively little is known about the biologic consequences of NOVA action and in particular about functional differences between NOVA1 and NOVA2.Thus we have discovered that NOVA2 uniquely regulates alternative splicing of a coordinate set of transcripts encoding key components in cortical, brainstem and spinal axon guidance/outgrowth pathways during neural differentiation, with severe functional consequences in vivo.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Neuro-Oncology, Howard Hughes Medical Institute, The Rockefeller University, New York, United States.

ABSTRACT
The neuron specific RNA-binding proteins NOVA1 and NOVA2 are highly homologous alternative splicing regulators. NOVA proteins regulate at least 700 alternative splicing events in vivo, yet relatively little is known about the biologic consequences of NOVA action and in particular about functional differences between NOVA1 and NOVA2. Transcriptome-wide searches for isoform-specific functions, using NOVA1 and NOVA2 specific HITS-CLIP and RNA-seq data from mouse cortex lacking either NOVA isoform, reveals that NOVA2 uniquely regulates alternative splicing events of a series of axon guidance related genes during cortical development. Corresponding axonal pathfinding defects were specific to NOVA2 deficiency: Nova2-/- but not Nova1-/- mice had agenesis of the corpus callosum, and axonal outgrowth defects specific to ventral motoneuron axons and efferent innervation of the cochlea. Thus we have discovered that NOVA2 uniquely regulates alternative splicing of a coordinate set of transcripts encoding key components in cortical, brainstem and spinal axon guidance/outgrowth pathways during neural differentiation, with severe functional consequences in vivo.

No MeSH data available.


Related in: MedlinePlus