Substrate Specificity and Cell Cycle Regulation of the Nek2 Protein Kinase, a Potential Human Homolog of the Mitotic Regulator NIMA of Aspergillus nidulans

Fry, Andrew M.; Schultz, Sharon J.; Bártek, Jiří; Nigg, Erich A.

doi:10.1074/jbc.270.21.12899

Cited by 141 publications

(142 citation statements)

References 23 publications

Supporting

Mentioning

135

Contrasting

Order By: Relevance

“…The Nek proteins belong to a family of serine/threonine kinases named after the NIMA gene originally cloned in Aspergillus nidulans (14). In mammals, there are 11 family members (Nek1-Nek11) but only four of these proteins, Nek2, Nek6, Nek7 and Nek9, play roles in mitotic progression (15).…”

Section: Introductionmentioning

confidence: 99%

Role of Nek2 on centrosome duplication and aneuploidy in breast cancer cells

et al. 2013

View full text Add to dashboard Cite

Breast cancer is the most common solid tumor and the second most common cause of death in women. Despite a large body of literature and progress in breast cancer research, many molecular aspects of this complex disease are still poorly understood, hindering the design of specific and effective therapeutic strategies. To identify molecules important in breast cancer progression and metastasis, we tested the in vivo effects of inhibiting the functions of various kinases and genes involved in the regulation/modulation of the cytoskeleton by downregulating them in mouse PyMT mammary tumor cells and human breast cancer cell lines. These kinases and cytoskeletal regulators were selected based on their prognostic values for breast cancer patient survival. PyMT tumor cells in which a selected gene was stably knocked down were injected into the tail veins of mice and the formation of tumors in the lungs was monitored. One of several genes found to be important for tumor growth in lungs was Nek2, a cell cycle-related protein kinase. Furthermore, Nek2 was also important for tumor growth in the mammary fat pad.In various human breast cancer cell lines, Nek2 knockdown induced aneuploidy and cell cycle arrest that led to cell death. Significantly, the breast cancer cell line most sensitive to Nek2 depletion was of the triple negative breast cancer subtype. Our data indicate that Nek2 plays a pivotal role in breast cancer growth at primary and secondary sites, and thus may be an attractive and novel therapeutic target for this disease.

show abstract

Section: Introductionmentioning

confidence: 99%

Role of Nek2 on centrosome duplication and aneuploidy in breast cancer cells

et al. 2013

View full text Add to dashboard Cite

show abstract

“…NEK2 is regulated through the cell cycle (Schultz et al, 1994;Fry et al, 1995) and has been localized to, and been shown to regulate, the centrosome (Fry et al, 1998b;Uto and Sagata, 2000) through interactions with the centrosomal protein C-nap1 (Fry et al, 1998a) and protein phosphatase type 1 (Helps et al, 2000). Additional roles for NEK2 during cell cycle progression are suggested by the localization of NEK2 to meiotic (Rhee and Wolgemuth, 1997) and mitotic chromosomes (Ha et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

TINA Interacts with the NIMA Kinase inAspergillus nidulansand Negatively Regulates Astral Microtubules during Metaphase Arrest

Osmani

Davies

Oakley

et al. 2003

MBoC

View full text Add to dashboard Cite

The tinA gene of Aspergillus nidulans encodes a protein that interacts with the NIMA mitotic protein kinase in a cell cycle-specific manner. Highly similar proteins are encoded in Neurospora crassa and Aspergillus fumigatus. TINA and NIMA preferentially interact in interphase and larger forms of TINA are generated during mitosis. Localization studies indicate that TINA is specifically localized to the spindle pole bodies only during mitosis in a microtubule-dependent manner. Deletion of tinA alone is not lethal but displays synthetic lethality in combination with the anaphase-promoting complex/cyclosome mutation bimE7. At the bimE7 metaphase arrest point, lack of TINA enhanced the nucleation of bundles of cytoplasmic microtubules from the spindle pole bodies. These microtubules interacted to form spindles joined in series via astral microtubules as revealed by live cell imaging. Because TINA is modified and localizes to the spindle pole bodies at mitosis, and lack of TINA causes enhanced production of cytoplasmic microtubules at metaphase arrest, we suggest TINA is involved in negative regulation of the astral microtubule organizing capacity of the spindle pole bodies during metaphase. INTRODUCTIONMitosis is regulated by cell cycle-specific phosphorylation and regulated proteolysis of regulatory proteins mediated by activation and inactivation of cyclin-dependent kinases (CDKs) (O'Farrell, 2001) in all eukaryotes, including Aspergillus nidulans . In A. nidulans, mitosis is also regulated by a second kinase, NIMA (never in mitosis A), which when inactivated arrests cell in G 2 even though the CDK1 complex is active (Osmani et al., 1991a). Without activation of NIMA, active CDK1 is unable to accumulate in the nucleus but does so after mutation of the nuclear pore complex protein SONA rae1/gle2 (Wu et al., 1998). NIMA may therefore interact with the nuclear pore complex to help mediate localization of CDK1 at mitosis.NIMA is regulated at multiple levels, including mRNA abundance (Osmani et al., 1987), phosphorylation , and proteolysis Ye et al., 1998). NIMA has also been shown to have a dynamic localization during mitosis being sequentially located to DNA, the mitotic spindle, and the spindle pole body (SPB) (De Souza et al., 2000).Expression of NIMA promotes chromatin condensation (O'Connell et al., 1994) and transient formation of mitotic spindle-like structures (Osmani et al., 1988b). Stable versions of NIMA prevent normal exit from mitosis . The mitotic promoting activity of NIMA crosses species barriers from yeast to humans (Lu and Hunter, 1995a), indicating conserved NIMA substrates are involved in mitotic regulation.NIMA-related kinases have been isolated from Neurospora crassa and Schizosaccharomyces pombe (Krien et al., 1998), and the S. pombe NIMA-like kinase Fin1p is involved in mitotic regulation (Grallert and Hagan, 2002;Krien et al., 2002). NIMA-related kinases (NEKs) have also been identified in higher eukaryotes (Letwin et al., 1992;Schultz and Nigg, 1993;Lu and Hunter, 1995b;Chen et al., 1999;Tana...

show abstract

“…The total digested plasmids were used to biolistically transform CU522 cells, and transformants were selected in SPP with 20 M paclitaxel. To prepare "kinase-dead" variants, plasmids with NRK1-GFP and NRK2-GFP coding regions were subjected to in vitro mutagenesis, to change the conserved lysine codon of the kinase domain to arginine at positions 40 and 35, respectively (Fry et al, 1995), using the QuikChange site-directed mutagenesis kit (Stratagene, La Jolla, CA). …”

mentioning

confidence: 99%

Members of the NIMA-related Kinase Family Promote Disassembly of Cilia by Multiple Mechanisms

Włoga

Camba

Rogowski

et al. 2006

MBoC

101

View full text Add to dashboard Cite

The genome of Tetrahymena thermophila contains 39 loci encoding NIMA-related kinases (NRKs), an extraordinarily large number for a unicellular organism. Evolutionary analyses grouped these sequences into several subfamilies, some of which have orthologues in animals, whereas others are protist specific. When overproduced, NRKs of three subfamilies caused rapid shortening of cilia. Ultrastructural studies revealed that each NRK triggered ciliary resorption by a distinct mechanism that involved preferential depolymerization of a subset of axonemal microtubules, at either the distal or proximal end. Overexpression of a kinase-inactive variant caused lengthening of cilia, indicating that constitutive NRK-mediated resorption regulates the length of cilia. Each NRK preferentially resorbed a distinct subset of cilia, depending on the location along the anteroposterior axis. We also show that normal Tetrahymena cells maintain unequal length cilia. We propose that ciliates used a large number of NRK paralogues to differentially regulate the length of specific subsets of cilia in the same cell. INTRODUCTIONHow the size of organelles is determined is an important and largely unanswered question. Cilia have emerged as a model organelle to study the mechanism of size regulation (Marshall, 2002). The size of an organelle is determined by the balance between assembly pathways that deliver structural components and disassembly pathways that remove components. In cilia, the intraflagellar transport (IFT), a bidirectional motility system that operates inside cilia (Rosenbaum and Witman, 2002;Scholey, 2003), plays a key role in length regulation. The anterograde IFT supplies structural subunits during assembly (Piperno et al., 1996;Qin et al., 2005) and maintains the axoneme after assembly (Brown et al., 1999). The retrograde IFT recycles the IFT machinery (Signor et al., 1999) and removes ciliary structural components that turn over (Qin et al., 2004). This turnover is the basis of the disassembly pathway (Stephens, 1997;Marshall and Rosenbaum, 2001;Song and Dentler, 2001;Thazhath et al., 2004). Experimental data and mathematical modeling in Chlamydomonas reinhardtii indicate that the length of flagella is dependent on the rates of elongation (equivalent to the efficiency of anterograde IFT) and disassembly. As the flagellum grows, the rate of elongation decreases due to the limited supply of IFT components that need to travel longer distances to the tip (Marshall and Rosenbaum, 2001). According to the "balance point model," the steady-state length is achieved when the rate of elongation equals the rate of disassembly (Marshall and Rosenbaum, 2001;Marshall et al., 2005).The length of already assembled cilia can be reduced drastically using two mechanisms: autotomy or resorption (reviewed by Quarmby, 2004). During autotomy, the axoneme breaks off from the basal body, the cilium falls off, and the plasma membrane reseals over the basal body. Protists undergo autotomy in response to low pH (Lewin et al., 1982). It is unclear whether...

show abstract

Substrate Specificity and Cell Cycle Regulation of the Nek2 Protein Kinase, a Potential Human Homolog of the Mitotic Regulator NIMA of Aspergillus nidulans

Cited by 141 publications

References 23 publications

Role of Nek2 on centrosome duplication and aneuploidy in breast cancer cells

Role of Nek2 on centrosome duplication and aneuploidy in breast cancer cells

TINA Interacts with the NIMA Kinase inAspergillus nidulansand Negatively Regulates Astral Microtubules during Metaphase Arrest

Members of the NIMA-related Kinase Family Promote Disassembly of Cilia by Multiple Mechanisms

Contact Info

Product

Resources

About