N-Acetyl-D-Glucosamine Kinase Interacts with NudC and Lis1 in Dynein Motor Complex and Promotes Cell Migration.

Md Ariful Islam,Diyah Fatimah Oktaviani,Il Soo Moon,Dae-Hyun Seog,Ho Jin Choi,Raju Dash,Syeda Ridita Sharif

doi:10.3390/ijms22010129

Md Ariful Islam, Diyah Fatimah Oktaviani + Show 5 more

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https://doi.org/10.3390/ijms22010129

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Abstract

Recently, we showed that N-acetylglucosamine kinase (NAGK), an enzyme of amino sugar metabolism, interacts with dynein light chain roadblock type 1 (DYNLRB1) and promotes the functions of dynein motor. Here, we report that NAGK interacts with nuclear distribution protein C (NudC) and lissencephaly 1 (Lis1) in the dynein complex. Yeast two-hybrid assays, pull-down assays, immunocytochemistry, and proximity ligation assays revealed NAGK–NudC–Lis1–dynein complexes around nuclei, at the leading poles of migrating HEK293T cells, and at the tips of migratory processes of cultured rat neuroblast cells. The exogenous expression of red fluorescent protein (RFP)-tagged NAGK accelerated HEK293T cell migration during in vitro wound-healing assays and of neurons during in vitro neurosphere migration and in utero electroporation assays, whereas NAGK knockdown by short hairpin RNA (shRNA) delayed migration. Finally, a small NAGK peptide derived from the NudC interacting domain in in silico molecular docking analysis retarded the migrations of HEK293T and SH-SY5Y cells. These data indicate a functional interaction between NAGK and dynein–NudC–Lis1 complex at the nuclear envelope is required for the regulation of cell migration.

Highlights

Cell migration takes place during diverse developmental and pathogenic processes in multicellular organisms
N-acetylglucosamine kinase (NAGK) interacts with the perinuclear Dynein–lissencephaly 1 (Lis1)–NudE1 complex during prophase nuclear invagination and with kinetochores during metaphase chromosome separation [24]. These findings suggest that NAGK plays an essential role in the molecular tug of war required for nuclear envelope breakdown and chromosome separation, during which the dynein motor complex is aided by bridging proteins such as NAGK, nuclear distribution protein C (NudC), and Lis1 [30]
The C-terminal contains a CSdomain (PFAM accession number: PF04969) [31]. This bipartite domain is of ≈100 residues, and its protein–protein interaction module is composed of a compact antiparallel β-sandwich fold consisting of seven β-strands

Summary

Introduction

Cell migration takes place during diverse developmental and pathogenic processes in multicellular organisms. Neuron migration during embryonic brain development essentially guides the complex layering of many structural and functional compartments. Neuronal progenitor cells (NPCs) are generated in spatially restricted ventricular zones (VZs) and undergo long-distance migration to reach their final destinations, e.g., the cortical plate (CP), to establish functional integrated neural circuitry [1,2]. Bipolar neurons extend a single or branched “migratory” leading process in the direction of movement. One or more transient “swellings” or “dilations” are formed within these leading processes that represent sites of attachment to underlying radial glial cells in vivo, and these dilations are observed in migratory bipolar cells of dissociated neuronal cultures in vitro and in other cases of non-glial guided migration [4,5]. Centrosomes move toward a swelling [6], and the cell body subsequently catches up with movement in a series of discontinuous steps, i.e., by nucleus–centrosome (N-C)

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