Discovering novel protein-protein interactions of Crk/Crkl adaptor proteins in retinal neurodevelopment

Conference Year

January 2021

Abstract

Nervous system development is a complex interplay of cell signaling and communication, with even slight variations having the potential to wreak havoc on typical development. CRK adaptor proteins (CRK and CRK-like, or CRKL), act to tether upstream signaling molecules such as membrane receptors with downstream signaling molecules to propagate cell signaling. They have an established role in embryonic eye development, proven through utilizing CRISPR-Cas9-generated mutant alleles, with loss of CRK and CRKL resulting in edema of the brain and heart, decreased eye size, shorter body axis, and defective retinal layering (Stergas et al., unpublished). Deletion of the 17p13 chromosome containing CRK results in a ‘smooth-brain syndrome’ known as Miller-Dieker syndrome, resulting in cognitive delays, cerebral palsy, and epilepsy (Hsieh, Jennesson, Thiele, Caruso, Masiakos & Duhaime, 2013). Deletion of chromosome 22q11.2 containing CRKL is associated with DiGeorge Syndrome, a neural crest migratory disease leading to pronounced craniofacial abnormalities and cleft palate, as well as impacting cardiovascular, immune, circulatory, and nervous systems (Formin, Pastorino, Kim, Pereira, Carneiro-Sampaio & Abe-Jacob, 2010). We seek to identify novel interacting partners of CRK/CRKL adaptor proteins to better characterize their role in retinal lamination during development and associated human disease states. We extract protein from zebrafish embryos and perform protein binding assays to identify what proteins zebrafish CRK adaptors are interacting with using biochemistry and mass spectrometry. This will provide the initial protein-protein interactions in the CRK/CRKL signaling pathway helping to elucidate the full signaling pathway of these important proteins.

Primary Faculty Mentor Name

Alicia Ebert

Graduate Student Mentors

Helaina Stergas

Faculty/Staff Collaborators

Helaina Stergas (Graduate Student Mentor)

Status

Undergraduate

Student College

College of Arts and Sciences

Program/Major

Neuroscience

Primary Research Category

Biological Sciences

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Discovering novel protein-protein interactions of Crk/Crkl adaptor proteins in retinal neurodevelopment

Nervous system development is a complex interplay of cell signaling and communication, with even slight variations having the potential to wreak havoc on typical development. CRK adaptor proteins (CRK and CRK-like, or CRKL), act to tether upstream signaling molecules such as membrane receptors with downstream signaling molecules to propagate cell signaling. They have an established role in embryonic eye development, proven through utilizing CRISPR-Cas9-generated mutant alleles, with loss of CRK and CRKL resulting in edema of the brain and heart, decreased eye size, shorter body axis, and defective retinal layering (Stergas et al., unpublished). Deletion of the 17p13 chromosome containing CRK results in a ‘smooth-brain syndrome’ known as Miller-Dieker syndrome, resulting in cognitive delays, cerebral palsy, and epilepsy (Hsieh, Jennesson, Thiele, Caruso, Masiakos & Duhaime, 2013). Deletion of chromosome 22q11.2 containing CRKL is associated with DiGeorge Syndrome, a neural crest migratory disease leading to pronounced craniofacial abnormalities and cleft palate, as well as impacting cardiovascular, immune, circulatory, and nervous systems (Formin, Pastorino, Kim, Pereira, Carneiro-Sampaio & Abe-Jacob, 2010). We seek to identify novel interacting partners of CRK/CRKL adaptor proteins to better characterize their role in retinal lamination during development and associated human disease states. We extract protein from zebrafish embryos and perform protein binding assays to identify what proteins zebrafish CRK adaptors are interacting with using biochemistry and mass spectrometry. This will provide the initial protein-protein interactions in the CRK/CRKL signaling pathway helping to elucidate the full signaling pathway of these important proteins.