| 1997 |
TLK1 (PKU-beta) is a serine/threonine protein kinase that localizes to the nucleus via an N-terminal nuclear localization signal; the NLS-containing GST-fusion was shown to localize to the nucleus in transfection experiments, and PKU-beta expressed in COS-1 cells was predominantly nuclear. |
Biochemical cloning, GST-fusion nuclear localization assay, transient transfection in COS-1 cells |
Gene |
Medium |
9427565
|
| 2003 |
TLK1 phosphorylates histone H3 in vitro and in vivo; expression of a kinase-dead TLK1B dominant negative mutant reduced phosphorylated histone H3 levels, caused chromosome condensation defects, chromosome missegregation, and aneuploidy in normal diploid breast epithelial cells. siRNA-mediated TLK1 depletion caused S-phase cell cycle block. |
Kinase-dead dominant negative overexpression, FACS, immunofluorescence microscopy, siRNA knockdown |
BMC cell biology |
High |
14583098
|
| 2005 |
TLK1 phosphorylates the DEAD-box RNA helicase p68 (DDX5); in vitro kinase assay showed phosphorylation of the p68 C-terminal fragment by recombinant TLK1, and overexpression of TLK1 in HT1080 cells increased phosphorylation of endogenous p68. Phosphorylation of the p68 C-terminal fragment by TLK1 reduced its affinity for single-stranded RNA. |
In vitro kinase assay, gel shift assay, TLK1 overexpression in HT1080 cells, immunoprecipitation |
Biochemical and biophysical research communications |
Medium |
15950181
|
| 2008 |
TLK1B binds Rad9 (component of the 9-1-1 complex) and phosphorylates hRad9 at S328; TLK1B overexpression hastened repair of DSBs, modulated 9-1-1 complex occupancy at DSBs, and stimulated chromatin reassembly adjacent to a DSB independently of kinase activity (kinase-dead TLK1B also stimulated chromatin assembly), suggesting a chaperone-like function. Rad9 and Asf1 compete for binding to TLK1B. |
Co-immunoprecipitation, in vitro kinase assay, S328A mutagenesis, HO endonuclease-induced DSB, chromatin assembly assay, chromatin immunoprecipitation |
DNA repair |
High |
18940270
|
| 2008 |
TLK1 (PKU-beta) depletion by RNAi caused reduced phosphorylation of myosin II regulatory light chain (MRLC) at Ser19/Thr18 and resulted in unequal chromosome segregation; expression of phosphomimetic DD-MRLC rescued chromosome segregation defects, indicating TLK1 regulates mitosis via MRLC phosphorylation. |
RNAi knockdown, immunoblot, immunofluorescence, rescue with DD-MRLC expression |
Mutation research |
Medium |
18838128
|
| 2010 |
In response to severe hypoxia, Chk1 is activated and phosphorylates TLK1, leading to its inactivation; this places TLK1 downstream of Chk1 in the DNA damage response pathway triggered by hypoxia-induced replication arrest. |
Cell-based assays under hypoxia, Chk1 loss-of-function, phosphorylation analysis |
Cell cycle (Georgetown, Tex.) |
Medium |
20581459
|
| 2019 |
TLK1B is upregulated following androgen deprivation therapy (ADT) and acts upstream of NEK1 and ATR to mediate the DNA damage response (DDR) in prostate cancer cells; the pathway ADT > TLK1 > NEK1 > ATR > Chk1 was established by showing that TLK1 inhibition with thioridazine impairs ATR and Chk1 activation and leads to apoptosis. |
TLK1 inhibitor (thioridazine) treatment, phosphorylation western blot, clonogenic survival assay, LNCaP xenografts, genetic NEK1-T141A mutant |
International journal of cancer |
High |
30737777
|
| 2019 |
TLK1 phosphorylates NEK1 at T141 to activate it; p-Nek1-T141 is elevated in prostate cancer biopsies correlating with Gleason score, and the TLK1-Nek1 axis is a common adaptive response during androgen-insensitive progression. |
In vitro phosphorylation, NEK1-T141A mutant xenografts, tissue microarray with pNek1-T141 antibody, TRAMP mouse model with TLK1 inhibitor |
Cancer letters |
High |
30928383
|
| 2020 |
TLK1 activates NEK1 via T141 phosphorylation, and this TLK1/NEK1 axis contributes to VDAC1 phosphorylation and stability, thereby maintaining mitochondrial permeability and integrity; cells overexpressing NEK1-T141A showed increased apoptosis, cytochrome C leakage, and altered mitochondrial metabolism upon DNA damage. |
NEK1-T141A mutant overexpression, doxorubicin treatment, oxygen consumption measurement, cytochrome C fractionation, cell cycle analysis |
Cell cycle (Georgetown, Tex.) |
Medium |
31914854
|
| 2020 |
In vitro kinase assays confirmed TLK1 inhibitory activity of the phenothiazine analog J54; J54 impairs the TLK1>NEK1>ATR>Chk1 DDR pathway and mediates apoptosis in prostate cancer cells and LNCaP xenografts. |
In vitro kinase assay, molecular docking, cell viability assays, xenograft model |
iScience |
Medium |
32905878
|
| 2021 |
TLK1 interacts with and phosphorylates AKTIP at T22 and S237; TLK1-mediated AKTIP phosphorylation enhances AKT association with PDK1 and promotes AKT phosphorylation at T308 and S473, connecting TLK1 to the PI3K/AKT survival pathway in prostate cancer. |
Interactome analysis, Co-IP, in vitro phosphorylation, AKTIP knockdown, phospho-western blot, TLK1 inhibitor J54 |
Pathophysiology |
Medium |
35366279
|
| 2022 |
TLK1 directly phosphorylates MK5 (MAPK-activated protein kinase 5) at S160, S354, and S386, resulting in MK5 activation; TLK1-mediated phosphorylation of MK5-S354 is required for prostate cancer cell motility, as MK5-S354A expression in MK5-null MEFs failed to rescue motility. This TLK1>MK5 axis drives PCa cell migration and invasion. |
In vitro kinase assay, phospho-specific antisera (pMK5-S354), Co-IP in HEK293 cells, MK5 knockout MEF rescue, 2D/3D motility assays, TLK1/MK5 KO and KD, TMA immunohistochemistry |
Molecular oncology |
High |
35064619
|
| 2023 |
TLK1 phosphorylates RAD54 at T41, T59 (N-terminal domain) and T700 (C-terminal domain); phosphorylation at T41 and T59 supports homologous recombination repair (HRR) and protects cells from DSB damage, while phosphorylation at T700 impairs HRR and reveals a new RAD54-CTD interaction with RAD51. |
ISce-I-GR-DsRed HRR reporter assay, TLK1 inhibition/depletion, site-directed mutagenesis, in vitro kinase assay, cytotoxicity assays, Co-IP with RAD51 |
Nucleic acids research |
High |
37439356
|
| 2024 |
TLK1 and TLK2 are hyper-autophosphorylated at their N-termini via homo- or hetero-dimerization; this hyper-autophosphorylation masks a conserved non-canonical PIP box and suppresses recruitment of TLK1/2 to damaged chromatin. Mutation of the PIP box abolishes recruitment to DNA damage sites, establishing that TLK1 associates with PCNA through its PIP box to reach damage sites. |
Autophosphorylation assay, PIP-box mutagenesis, laser micro-irradiation/live-cell imaging at damage sites, dimerization analysis, biochemical fractionation |
Nucleic acids research |
High |
39727191
|
| 2024 |
TLK1 (via NEK1) promotes nuclear retention and transcriptional activation of YAP through stabilizing phosphorylation at Y407; pYAP-Y407 shows stronger association with co-activators AR and TEAD1 and ChIP confirms YAP-wt but not YAP-Y407F occupancy at ARE- and TEAD1-driven gene promoters. TLK1 inhibitor J54 reverses YAP nuclear retention and dephosphorylates Y407. |
ChIP, nuclear/cytoplasmic fractionation, co-immunoprecipitation with AR/TEAD1, GFP-YAP-Y407F mutagenesis, TLK1 inhibitor J54 treatment, LNCaP and VCaP xenografts |
Cancers |
Medium |
39199688
|
| 2024 |
A TLK1 variant (p.Q479E) associated with a neurodevelopmental disorder impairs kinase activity but does not strongly alter TLK1 localization or proximal protein interactions; cells expressing this variant showed elevated spontaneous DNA damage and reduced cytokine responses, indicating TLK1 kinase activity is required for genomic and immune homeostasis. |
Patient-derived lymphoblast cell lines, single-cell gel electrophoresis (comet assay), western blot for DNA damage markers, flow cytometry, RNA-seq, biochemical kinase activity assay |
iScience |
Medium |
38868186
|