TNK1 is a ubiquitin-binding and 14-3-3-regulated kinase that can be targeted to block tumor growth.

TNK1 is a non-receptor tyrosine kinase with poorly understood biological function and regulation. Here, we identify TNK1 dependencies in primary human cancers. We also discover a MARK-mediated phosphorylation on TNK1 at S502 that promotes an interaction between TNK1 and 14-3-3, which sequesters TNK1 and inhibits its kinase activity. Conversely, the release of TNK1 from 14-3-3 allows TNK1 to cluster in ubiquitin-rich puncta and become active. Active TNK1 induces growth factor-independent proliferation of lymphoid cells in cell culture and mouse models. One unusual feature of TNK1 is a ubiquitin-association domain (UBA) on its C-terminus. Here, we characterize the TNK1 UBA, which has high affinity for poly-ubiquitin. Point mutations that disrupt ubiquitin binding inhibit TNK1 activity. These data suggest a mechanism in which TNK1 toggles between 14-3-3-bound (inactive) and ubiquitin-bound (active) states. Finally, we identify a TNK1 inhibitor, TP-5801, which shows nanomolar potency against TNK1-transformed cells and suppresses tumor growth in vivo.

Epithelial cell extrusion requires the sphingosine-1-phosphate receptor 2 pathway.

To maintain an intact barrier, epithelia eliminate dying cells by extrusion. During extrusion, a cell destined for apoptosis signals its neighboring cells to form and contract a ring of actin and myosin, which squeezes the dying cell out of the epithelium. Here, we demonstrate that the signal produced by dying cells to initiate this process is sphingosine-1-phosphate (S1P). Decreasing S1P synthesis by inhibiting sphingosine kinase activity or by blocking extracellular S1P access to its receptor prevented apoptotic cell extrusion. Extracellular S1P activates extrusion by binding the S1P(2) receptor in the cells neighboring a dying cell, as S1P(2) knockdown in these cells or its loss in a zebrafish mutant disrupted cell extrusion. Because live cells can also be extruded, we predict that this S1P pathway may also be important for driving delamination of stem cells during differentiation or invasion of cancer cells.

Ontogeny of salinity tolerance and evidence for seawater-entry preparation in juvenile green sturgeon, Acipenser medirostris.

We measured the ontogeny of salinity tolerance and the preparatory hypo-osmoregulatory physiological changes for seawater entry in green sturgeon (Acipenser medirostris), an anadromous species occurring along the Pacific Coast of North America. Salinity tolerance was measured every 2 weeks starting in 40-day post-hatch (dph) juveniles and was repeated until 100% survival at 34‰ was achieved. Fish were subjected to step increases in salinity (5‰ 12 h(-1)) that culminated in a 72-h exposure to a target salinity, and treatment groups (0, 15, 20, 25, 30, 34‰; and abrupt exposure to 34‰) were adjusted as fish developed. After 100% survival was achieved (134 dph), a second experiment tested two sizes of fish for 28-day seawater (33‰) tolerance, and gill and gastrointestinal tract tissues were sampled. Their salinity tolerance increased and plasma osmolality decreased with increasing size and age, and electron microscopy revealed three types of mitochondria-rich cells: one in fresh water and two in seawater. In addition, fish held on a natural photoperiod in fresh water at 19°C showed peaks in cortisol, thyroid hormones and gill and pyloric ceca Na(+), K(+)-ATPase activities at body sizes associated with seawater tolerance. Therefore, salinity tolerance in green sturgeon increases during ontogeny (e.g., as these juveniles may move down estuaries to the ocean) with increases in body size. Also, physiological and morphological changes associated with seawater readiness increased in freshwater-reared juveniles and peaked at their seawater-tolerant ages and body sizes. Their seawater-ready body size also matched that described for swimming performance decreases, presumably associated with downstream movements. Therefore, juvenile green sturgeon develop structures and physiological changes appropriate for seawater entry while growing in fresh water, indicating that hypo-osmoregulatory changes may proceed by multiple routes in sturgeons.