Alternative titles; symbolsNUDE-LIKE PROTEIN; NUDELENDOOLIGOPEPTIDASE A; EOPAHGNC Approved Gene Symbol: NDEL1Cytogenetic location: 17p13.1 Genomic coordinate...
Alternative titles; symbols
HGNC Approved Gene Symbol: NDEL1
Cytogenetic location: 17p13.1 Genomic coordinates (GRCh38): 17:8,435,883-8,472,743 (from NCBI)
▼ Cloning and Expression
Using mouse Lis1 (PAFAH1B1; 601545) as bait in a yeast 2-hybrid screen, Niethammer et al. (2000) cloned NUDEL from a human fetal brain cDNA library. The deduced protein contains 345 amino acids and has a calculated molecular mass of 38.4 kD. It has a coiled-coil motif (residues 19 to 201), followed by several potential phosphorylation sites for casein kinase II (see 115440), protein kinase C (see 176960), or CDK5 (123831). NUDEL shares about 50% identity with mouse and human NUDE proteins. Western blot analysis of mouse tissues showed abundant expression of Nudel in brain and testis and much lower expression in heart, liver, kidney, and skeletal muscle. In fractionated rat brain, Nudel and Lis1 were both found in fractions enriched for postsynaptic density proteins. Immunostaining of embryonic day-18 mouse brain sections revealed staining of migrating neurons and thalamocortical axons of the intermediate zone of the developing cerebral cortex, as well as several other developing brain regions. In COS-7 cells, endogenous Lis1 and Nudel colocalized at centrosome-like structures near the nucleus and were prominent at the center of the interphase microtubule array. In embryonic rat hippocampal neurons, Nudel was prominent at centrosomes and growth cones.
Using Western blot analysis, Yan et al. (2003) detected NUDEL at an apparent molecular mass of about 40 kD in several human cell lines. Western blot analysis of mouse tissues detected highest expression in brain, with much lower expression in heart, skeletal muscle, and lung, and little to no expression in other tissues examined.
Hayashi et al. (2005) cloned NUDEL from a human temporal cortex cDNA library. They determined that NUDEL is identical to endooligopeptidase A, a thiol-activated peptidase involved in the conversion or inactivation of a number of bioactive peptides.
▼ Gene Function
Using a yeast 2-hybrid assay, Niethammer et al. (2000) found that only full-length LIS1 was able to interact with NUDEL. They also coimmunoprecipitated LIS1 and NUDEL from COS-7 cells exogenously expressing both proteins and from mouse brain lysates. In GST pull-down experiments of rat brain extracts or purified protein preparations, GST-NUDEL interacted with cytoplasmic dynein intermediate chains, heavy chains, and light-intermediate chains (see 600112). Overexpression of mouse Lis1 in COS-7 cells caused a redistribution of transfected NUDEL. Biochemical and mutation analyses revealed that NUDEL is phosphorylated on 3 sites by CDK5. Overexpression of a phosphorylation-minus NUDEL mutant or inhibition of CDK5 produced neuritic swellings in rat brain cortical neurons.
Yan et al. (2003) found that NUDEL was phosphorylated on its serine/threonine phosphorylation motifs in M phase of the cell cycle in human cells. A fraction of NUDEL bound strongly to centrosomes in interphase and localized to mitotic spindles in early M phase. Using phosphorylation-deficient NUDEL mutants, Yan et al. (2003) confirmed that phosphorylation regulated the cell cycle-dependent distribution of NUDEL. Moreover, phosphorylated NUDEL or a phosphorylation-mimicking mutant bound PAFAH1B1 more efficiently. A mutant incapable of binding to PAFAH1B1 impaired the poleward movement of dynein and dynein-mediated transport of kinetochore proteins to spindle poles along microtubules, thus contributing to inactivation of the spindle checkpoint in mitosis. Yan et al. (2003) concluded that the interaction between NUDEL and PAFAH1B1 is important in regulating dynein activity in M phase.
Using a yeast 2-hybrid assay, Ozeki et al. (2003) found that DISC1 (605210) interacts with human brain NUDEL. They also found that the schizophrenia-associated form of DISC1, which has a 257-residue truncation at its C terminus, does not interact with NUDEL.
By analysis of crystalline structures of mouse proteins, Tarricone et al. (2004) determined that a Lis1 homodimer binds with a homodimer of either Pafah1b2 (602508) or Ndel1 to form a tetramer. Ndel1 competes with the Pafah1b2 homodimer for Lis1, but the interaction is complex and requires both the N- and C-terminal domains of Lis1. The data suggested that the Lis1 molecule undergoes major conformational changes when switching from a complex with the acetylhydrolase to that with Ndel1.
Hayashi et al. (2005) demonstrated that recombinant NUDEL could hydrolyze biologically active peptides of various sizes and sequences. NUDEL sowed broad substrate specificity, cleaving the phe5-ser6 bond of bradykinin (see 113503), the leu5-arg6 bond of dynorphin A (131340), and the arg9-arg10 bond of neurotensin (162650). Activity was enhanced by dithiolthretol and inhibited by thiol-reactive compounds. Mutation of the catalytic cys273 of NUDEL to alanine inactivated the enzyme. Gel-filtration chromatography of rat brain cytosol indicated that the active protein is monomeric. Hayashi et al. (2005) noted that the catalytic cysteine of NUDEL is close to the DISC1-binding site, and they demonstrated that DISC1 is a noncompetitive inhibitor of recombinant NUDEL oligopeptidase activity in vitro.
Using RNA interference (RNAi) with cultured cell lines and mouse embryonic day-15 cortical neurons, Shu et al. (2004) determined that Ndel1 regulates dynein activity by facilitating the interaction between Lis1 and dynein. Loss of Ndel1, Lis1, or dynein function in developing neocortex impaired neuronal positioning and caused the uncoupling of centrosomes and nuclei. Overexpression of Lis1 partially rescued the positioning defect caused by Ndel1 RNAi but not that caused by dynein RNAi, whereas overexpression of Ndel1 did not rescue the phenotype induced by Lis1 RNAi. Shu et al. (2004) concluded that NDEL1 interacts with LIS1 to sustain the function of dynein, which in turn impacts microtubule organization, nuclear translocation, and neuronal positioning.
Toyo-Oka et al. (2005) identified KATNA1 (606696) the microtubule-severing protein as a molecular target of NDEL1. Phosphorylation of Ndel1 by Cdk5 (123831) facilitated interaction between Ndel1 and Katna1, suggesting that phosphorylated NDEL1 may regulate the distribution of KATNA1. Abnormal accumulation of Katna1 in the nucleus of Ndel1-null mouse embryonic fibroblasts supported an essential role for NDEL1 in KATNA1 regulation. Complete loss of NDEL1 or expression of dominant-negative Katna1 mutants in migrating neurons resulted in defective migration and elongation of nuclear-centrosomal distance. Toyo-Oka et al. (2005) suggested that NDEL1 may be essential for mitotic cell division and neuronal migration, not only via regulation of cytoplasmic dynein function, but also by modulation of KATNA1 localization and function.
Shen et al. (2008) showed that Nudel colocalized with Cdc42gap (ARHGAP1; 602732) at the leading edge of migrating NIH3T3 mouse fibroblasts. This localization of Nudel required its phosphorylation by Erk1 (MAPK3; 601795)/Erk2 (MAPK1; 176948). Shen et al. (2008) found that Nudel competed with Cdc42 (116952) for binding Cdc42gap. Consequently, Nudel inhibited Cdc42gap-mediated inactivation of Cdc42 in a dose-dependent manner. Depletion of Nudel by RNA interference or overexpression of a nonphosphorylatable Nudel mutant abolished Cdc42 activation and cell migration. Shen et al. (2008) concluded that NUDEL facilitates cell migration by sequestering CDC42GAP at the leading edge to stabilize active CDC42 in response to extracellular stimuli.
Burdick et al. (2008) noted that NDE1 (609449) is a homolog of NDEL1 and also binds to DISC1. NDE1 was expressed at constant levels in the rat cerebral cortex from embryonic day (E) 14 to adulthood, whereas NDEL1 expression showed a time-course increase peaking at postnatal day 7. Further studies with a ser704-to-cys (S704C) polymorphism in the DISC1 gene showed that NDE1 bound stronger to ser704, while NDEL1 bound stronger to cys704. The findings suggested an interaction of these 3 proteins, with possible competitive binding between NDEL1 and NDE1 for DISC1.
Using cultured mouse neurons, Mori et al. (2009) showed that atypical protein kinase C (aPKC; see 176982), Aurora A (AURKA; 603072), and Ndel1 function in a pathway that regulates microtubule organization during neurite extension. aPKC threonine phosphorylated Aurora A, which augmented interaction with Tpx2 (605917) and facilitated activation of Aurora A at the neurite hillock. Aurora A then serine phosphorylated and recruited Ndel1. Suppression of aPKC, Aurora A, or Tpx2, or disruption of Ndel1, severely impaired neurite extension, and this impairment was accompanied by reduced frequency of microtubule emanation from the microtubule organizing center. Mori et al. (2009) concluded that aPKC, Aurora A, and NDEL1 comprise a signaling pathway for microtubule remodeling during neurite extension.
The International Radiation Hybrid Mapping Consortium mapped the NUDEL gene to chromosome 17 (sts-N53016).