The Dyslexia-associated KIAA0319 Protein Undergoes Proteolytic Processing with γ-Secretase-independent Intramembrane Cleavage

The KIAA0319 gene has been associated with reading disability in several studies. It encodes a plasma membrane protein with a large, highly glycosylated, extracellular domain. This protein is proposed to function in adhesion and attachment and thought to play an important role during neuronal migration in the developing brain. We have previously proposed that endocytosis of this protein could constitute an important mechanism to regulate its function. Here we show that KIAA0319 undergoes ectodomain shedding and intramembrane cleavage. At least five different cleavage events occur, four in the extracellular domain and one within the transmembrane domain. The ectodomain shedding processing cleaves the extracellular domain, generating several small fragments, including the N-terminal region with the Cys-rich MANEC domain. It is possible that these fragments are released to the extracellular medium and trigger cellular responses. The intramembrane cleavage releases the intracellular domain from its membrane attachment. Our results suggest that this cleavage event is not carried out by γ-secretase, the enzyme complex involved in similar processing in many other type I proteins. The soluble cytoplasmic domain of KIAA0319 is able to translocate to the nucleus, accumulating in nucleoli after overexpression. This fragment has an unknown role, although it could be involved in regulation of gene expression. The absence of DNA-interacting motifs indicates that such a function would most probably be mediated through interaction with other proteins, not by direct DNA binding. These results suggest that KIAA0319 not only has a direct role in neuronal migration but may also have additional signaling functions.

[1]  A. Monaco,et al.  The genetic lexicon of dyslexia. , 2007, Annual review of genomics and human genetics.

[2]  J. Stockley,et al.  Understanding BACE1: essential protease for amyloid-β production in Alzheimer’s disease , 2008, Cellular and Molecular Life Sciences.

[3]  Richard Wade-Martins,et al.  A Common Variant Associated with Dyslexia Reduces Expression of the KIAA0319 Gene , 2009, PLoS genetics.

[4]  Richard Wade-Martins,et al.  The chromosome 6p22 haplotype associated with dyslexia reduces the expression of KIAA0319, a novel gene involved in neuronal migration. , 2006, Human molecular genetics.

[5]  P. Saftig,et al.  Breaking up the tie: disintegrin-like metalloproteinases as regulators of cell migration in inflammation and invasion. , 2006, Pharmacology & therapeutics.

[6]  F. Murakami,et al.  The role of Slit-Robo signaling in the generation, migration and morphological differentiation of cortical interneurons. , 2008, Developmental biology.

[7]  Anthony P. Monaco,et al.  The dyslexia-associated protein KIAA0319 interacts with adaptor protein 2 and follows the classical clathrin-mediated endocytosis pathway , 2009, American journal of physiology. Cell physiology.

[8]  J. Arribas,et al.  Protein ectodomain shedding. , 2002, Chemical reviews.

[9]  H. Inoue,et al.  Membrane‐anchored growth factors, the epidermal growth factor family: Beyond receptor ligands , 2008, Cancer science.

[10]  Y. Kwon,et al.  The mammalian N-end rule pathway: new insights into its components and physiological roles. , 2007, Trends in biochemical sciences.

[11]  B. de Strooper,et al.  γ-Secretase activity requires the presenilin-dependent trafficking of nicastrin through the Golgi apparatus but not its complex glycosylation , 2003, Journal of Cell Science.

[12]  C. Francks,et al.  Genes, cognition and dyslexia: learning to read the genome , 2006, Trends in Cognitive Sciences.

[13]  M. Wolfe.  Intramembrane-cleaving Proteases* , 2009, Journal of Biological Chemistry.

[14]  B. de Strooper,et al.  Structure and function of gamma-secretase. , 2009, Seminars in cell & developmental biology.

[15]  Narmada Thanki,et al.  CDD: specific functional annotation with the Conserved Domain Database , 2008, Nucleic Acids Res..

[16]  G. Murphy.  The ADAMs: signalling scissors in the tumour microenvironment , 2008, Nature Reviews Cancer.

[17]  S. Hébert,et al.  Differential contribution of the three Aph1 genes to γ-secretase activity in vivo , 2005 .

[18]  C. Sanders,et al.  Substrate specificity of γ-secretase and other intramembrane proteases , 2008, Cellular and Molecular Life Sciences.

[19]  E. Wolf,et al.  The epidermal growth factor receptor ligands at a glance , 2009, Journal of cellular physiology.

[20]  C. Blobel,et al.  ADAMs: key components in EGFR signalling and development , 2005, Nature Reviews Molecular Cell Biology.

[21]  R. Vassar,et al.  BACE1 structure and function in health and Alzheimer's disease. , 2008, Current Alzheimer research.

[22]  H. Vanderstichele,et al.  Presenilin 2 deficiency causes a mild pulmonary phenotype and no changes in amyloid precursor protein processing but enhances the embryonic lethal phenotype of presenilin 1 deficiency. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[23]  P. Bork,et al.  Epidermal growth factor-like modules , 1993 .

[24]  A. Monaco,et al.  The dyslexia-associated gene KIAA0319 encodes highly N- and O-glycosylated plasma membrane and secreted isoforms. , 2008, Human molecular genetics.

[25]  A. Monaco,et al.  Alternative splicing in the dyslexia-associated gene KIAA0319 , 2007, Mammalian Genome.

[26]  M. Freeman,et al.  Intramembrane proteolysis controls diverse signalling pathways throughout evolution. , 2002, Current opinion in genetics & development.

[27]  Jeffrey R. Gruen,et al.  Progress towards a cellular neurobiology of reading disability , 2010, Neurobiology of Disease.

[28]  Long Yu,et al.  MANSC: a seven-cysteine-containing domain present in animal membrane and extracellular proteins. , 2004, Trends in biochemical sciences.

[29]  Ryan E. Mills,et al.  Classical Nuclear Localization Signals: Definition, Function, and Interaction with Importin α* , 2007, Journal of Biological Chemistry.

[30]  P. Dempsey,et al.  Control of ErbB signaling through metalloprotease mediated ectodomain shedding of EGF-like factors , 2006, Growth factors.

[31]  M. Pelto-huikko,et al.  Shedding light on ADAM metalloproteinases. , 2005, Trends in biochemical sciences.

[32]  Raphael Kopan,et al.  Presenilin: RIP and beyond. , 2009, Seminars in cell & developmental biology.

[33]  Joseph L Goldstein,et al.  Regulated Intramembrane Proteolysis A Control Mechanism Conserved from Bacteria to Humans , 2000, Cell.

[34]  Weixian Lu,et al.  A time- and cost-efficient system for high-level protein production in mammalian cells. , 2006, Acta crystallographica. Section D, Biological crystallography.

[35]  K. Klinger,et al.  Strong homophilic interactions of the Ig-like domains of polycystin-1, the protein product of an autosomal dominant polycystic kidney disease gene, PKD1. , 2000, Human molecular genetics.

[36]  B. de Strooper,et al.  A cell biological perspective on Alzheimer's disease. , 2002, Annual review of cell and developmental biology.

[37]  Lina L. Feng,et al.  Evolution of distinct EGF domains with specific functions , 2005, Protein science : a publication of the Protein Society.

[38]  L. Matrisian,et al.  Matrix metalloproteinases: they're not just for matrix anymore! , 2001, Current opinion in cell biology.

[39]  K. Inouye,et al.  Roles of Functional and Structural Domains of Hepatocyte Growth Factor Activator Inhibitor Type 1 in the Inhibition of Matriptase* , 2008, Journal of Biological Chemistry.

[40]  Marc Tessier-Lavigne,et al.  Roundabout Controls Axon Crossing of the CNS Midline and Defines a Novel Subfamily of Evolutionarily Conserved Guidance Receptors , 1998, Cell.

[41]  Marc P. Stemmler.  Cadherins in development and cancer. , 2008, Molecular bioSystems.

[42]  Bernd Bukau,et al.  The N-end rule pathway for regulated proteolysis: prokaryotic and eukaryotic strategies. , 2007, Trends in cell biology.

[43]  B. Pennington,et al.  Breakthroughs in the search for dyslexia candidate genes. , 2006, Trends in molecular medicine.

[44]  B. de Strooper,et al.  Presenilins: members of the gamma-secretase quartets, but part-time soloists too. , 2008, Physiology.

[45]  B. de Strooper,et al.  ADAM10, the Rate-limiting Protease of Regulated Intramembrane Proteolysis of Notch and Other Proteins, Is Processed by ADAMS-9, ADAMS-15, and the γ-Secretase* , 2009, Journal of Biological Chemistry.

[46]  F. Ramus,et al.  From genes to behavior in developmental dyslexia , 2006, Nature Neuroscience.

[47]  Raphael Kopan,et al.  γ-Secretase: proteasome of the membrane? , 2004, Nature Reviews Molecular Cell Biology.

[48]  D. Selkoe,et al.  Notch and Presenilin: regulated intramembrane proteolysis links development and degeneration. , 2003, Annual review of neuroscience.

[49]  K. Miyazawa,et al.  Hepatocyte Growth Factor Activator Inhibitor Type 1 Is a Specific Cell Surface Binding Protein of Hepatocyte Growth Factor Activator (HGFA) and Regulates HGFA Activity in the Pericellular Microenvironment* , 2000, The Journal of Biological Chemistry.

[50]  P. Saftig,et al.  The "a disintegrin and metalloprotease" (ADAM) family of sheddases: physiological and cellular functions. , 2009, Seminars in cell & developmental biology.

[51]  D. Seals,et al.  The ADAMs family of metalloproteases: multidomain proteins with multiple functions. , 2003, Genes & development.

[52]  B. Martoglio,et al.  Intramembrane-cleaving proteases: controlled liberation of proteins and bioactive peptides. , 2003, Trends in cell biology.

[53]  A. Galaburda,et al.  The effect of variation in expression of the candidate dyslexia susceptibility gene homolog Kiaa0319 on neuronal migration and dendritic morphology in the rat. , 2010, Cerebral cortex.

[54]  F. Boisvert,et al.  The multifunctional nucleolus , 2007, Nature Reviews Molecular Cell Biology.