N-Acetyl-D-Glucosamine Kinase Interacts with Dynein-Lis1-NudE1 Complex and Regulates Cell Division

Abstract

N-acetyl-D-glucosamine kinase (GlcNAc kinase or NAGK) primarily catalyzes phosphoryl transfer to GlcNAc during amino sugar metabolism. Recently, it was shown NAGK interacts with dynein light chain roadblock type 1 (DYNLRB1) and upregulates axo-dendritic growth, which is an enzyme activity-independent, non-canonical structural role. The authors examined the distributions of NAGK and NAGK-dynein complexes during the cell cycle in HEK293T cells. NAGK was expressed throughout different stages of cell division and immunocytochemistry (ICC) showed NAGK was localized at nuclear envelope, spindle microtubules (MTs), and kinetochores (KTs). A proximity ligation assay (PLA) for NAGK and DYNLRB1 revealed NAGK-dynein complex on nuclear envelopes in prophase cells and on chromosomes in metaphase cells. NAGK-DYNLRB1 PLA followed by Lis1/NudE1 immunostaining showed NAGK-dynein complexes were colocalized with Lis1 and NudE1 signals, and PLA for NAGK-Lis1 showed similar signal patterns, suggesting a functional link between NAGK and dynein-Lis1 complex. Subsequently, NAGK-dynein complexes were found in KTs and on nuclear membranes where KTs were marked with CENP-B ICC and nuclear membrane with lamin ICC. Furthermore, knockdown of NAGK by small hairpin (sh) RNA was found to delay cell division. These results indicate that the NAGK-dynein interaction with the involvements of Lis1 and NudE1 plays an important role in prophase nuclear envelope breakdown (NEB) and metaphase MT-KT attachment during eukaryotic cell division.

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

“…Human NAGK monomer is 37 kDa sized, composed of N-terminal (small) and C-terminal (large) domains [ 21 ], belongs to the sugar kinase/heat shock protein 70/actin superfamily [ 22 ], and is characterized by a V-shaped fold consisting of two domains. Findings from our laboratory and other research group suggest a critical role of NAGK in the early development process [ 23 , 24 , 25 ]. NAGK is expressed at high levels in neurons but at low levels in astrocytes and oligodendrocytes [ 26 , 27 ], and it plays essential roles during the development of dendrites [ 26 , 27 ] and axons [ 25 ], in protein aggregates clearance [ 28 ], and during cell division [ 24 ].…”

“…Findings from our laboratory and other research group suggest a critical role of NAGK in the early development process [ 23 , 24 , 25 ]. NAGK is expressed at high levels in neurons but at low levels in astrocytes and oligodendrocytes [ 26 , 27 ], and it plays essential roles during the development of dendrites [ 26 , 27 ] and axons [ 25 ], in protein aggregates clearance [ 28 ], and during cell division [ 24 ]. NAGK interacted with dynein light chain roadblock 1 (DYNLRB1) during yeast two-hybrid selection [ 29 ].…”

N-acetyl-D-glucosamine kinase binds dynein light chain roadblock 1 and promotes protein aggregate clearance

“…We favor the latter scenario for two reasons. First, NAGK promotes the function of cytoplasmic dynein [11][12][13][14][15] . If aggregates were disposed of via the ALP, efficient transport of aggregates to MTOC by dynein-NAGK complex would facilitate formation of aggresome, autophagosome, and autolysosome, because lysosomes are highly concentrated at the cell center (i.e., MTOC) 7 , accelerated transport would aid aggregate clearance.…”

“…In this regard, N -acetylglucosamine kinase (NAGK) is a good candidate for promoting dynein activation. Specifically, NAGK interacts with dynein light chain roadblock type 1 (DYNLRB1) 11 to promote dynein functions during mitosis 12 and axonal 13 and dendritic growth 14 , 15 . In the present study, we asked whether NAGK also promotes the clearance of protein aggregates.…”

Serine/Threonine Protein Kinase STK16

“…It is highly expressed in neurons [93], and the interaction of GlcNAcK-Dynein-Golgi could regulate the growth of axons [94] and dendrites [95] of neurons. Moreover, the interaction of GlcNAcK-Dynein-Lis1-NudE1 plays an important role in the prophase and metaphase of mitosis [96]. Meanwhile, GlcNAcK also has different subnuclear distribution [97].…”