Biology subjects

Lacigova, A.

Publications and source records attributed to Lacigova, A..

3 recordsLinked to original sources

bioRxiv2026

An Analog-Sensitive Tau Tubulin Kinase 2 Enables Acute Chemical-Genetic Control of Primary Cilia

Kinases regulate a broad spectrum of cellular processes, including the formation of primary cilia, microtubule-based organelles that protrude from the cell surface and play important roles in development and disease. TTBK2 is a key ciliary kinase that controls primary cilium assembly and maintenance. However, highly selective small-molecule inhibitors of TTBK2 are still lacking, representing a major bottleneck for investigating the temporal requirements and dynamics of TTBK2-dependent processes in cilia. Here, we report the first analog-sensitive variant of TTBK2, enabling rapid, reversible, and highly specific control of TTBK2 kinase activity in ciliogenesis-relevant hTERT-RPE1 cells. Using this system, we define the kinetics of CEP164 phosphorylation, cilium shortening and reassembly, and IFT88 retention at the ciliary base following acute TTBK2 inhibition and subsequent inhibitor washout. Together, our findings provide new insight into TTBK2-dependent pathways and establish analog-sensitive TTBK2 as a powerful tool for investigating the dynamics of TTBK2-regulated processes with high temporal resolution.

cell biology↗

bioRxiv2026

Development of Potent and Cell Active 5-Azaindole-Based Tau Tubulin Kinase Inhibitors

We have developed and characterized a potent and cell active tau tubulin kinase 1 and 2 (TTBK1 and TTBK2) inhibitor, 13. Compound 13 demonstrates in-cell, kinome-wide selectivity, and potently inhibits both TTBK1 and TTBK2. As part of our medicinal chemistry campaign, we also identified a structurally similar negative control, compound 5, which lacks in-cell affinity for TTBK1 and TTBK2. Based on their substrates, which include TDP-43, tau, and tubulin, TTBK1 and TTBK2 inhibition has been pursued as a therapeutic approach for Alzheimers disease, frontotemporal lobe dementia, and amyotrophic lateral sclerosis. TTBK2 is also an effector of ciliogenesis, acting in concert with CEP164, CP110, and CEP83 to initiate the biogenesis of primary cilia. The development of selective chemical tools for these kinases facilitates investigation into TTBK1/2-mediated pathways and potential disease-altering ramifications linked to their pharmacological perturbation.

cell biology↗

bioRxiv2023

Tau-Tubulin Kinase 2 restrains microtubule-depolymerizer KIF2A to support primary cilia growth.

The initiation of assembly of primary cilia, organelles with crucial functions in development and disease, is under the control of Tau-Tubulin Kinase 2 (TTBK2). Recent work has implicated TTBK2 also in the regulation of primary cilia maintenance and function. However, the mechanisms underlying individual functions of TTBK2 in primary cilia are not fully understood. Here, to dissect the role of TTBK2 in primary cilia maintenance in human cells, we examined disease related TTBK2 truncations. We demonstrate that these truncated protein moieties show selective activity towards TTBK2 substrates. This creates a semi-permissive condition where partial TTBK2 activity suffices to support the initiation of ciliogenesis but fails to sustain primary cilia length. Subsequently, we show that the defects in primary cilia growth are linked to aberrant turnover of kinesin KIF2A at basal body. Furthermore, we demonstrate that TTBK2 regulates KIF2A by phosphorylation, which in turn restrains its levels at the ciliary base to promote primary cilia elongation and maintenance. Taken together, our data highlight the regulation of KIF2A by TTBK2 as an important mechanism governing primary cilia in human cells.

cell biology↗