The characterisation of LATS2 kinase regulation in Hippo-YAP signalling

• Published in: Cellular signalling Vol. 28; no. 5; pp. 488 - 497
• Main Authors: Hoa, Lily, Kulaberoglu, Yavuz, Gundogdu, Ramazan, Cook, Dorthe, Mavis, Merdiye, Gomez, Marta, Gomez, Valenti, Hergovich, Alexander
• Format: Journal Article
• Language: English
• Published: England Elsevier Inc 01-05-2016
• Subjects: Adaptor Proteins, Signal Transducing - metabolism; Animals; Binding; Chlorocebus aethiops; Complex formation; Control; COS Cells; Gain; Government regulations; HEK293 Cells; Hippo-YAP tumour suppressor pathway; Humans; Intracellular kinase signalling; Kinases; Large tumour suppressor kinase; MOB proteins; Mutation; Phosphoproteins - metabolism; Phosphorylation; Protein-Serine-Threonine Kinases - chemistry; Protein-Serine-Threonine Kinases - genetics; Protein-Serine-Threonine Kinases - metabolism; Signal Transduction; Signalling; STK3/4; Transcription Factors; Tumor Suppressor Proteins - chemistry; Tumor Suppressor Proteins - genetics; Tumor Suppressor Proteins - metabolism; YAP1; Intracellular kinase signalling; MOB; HM; STK3/4; STK; PIF; PRK2; NTR; Hippo-YAP tumour suppressor pathway; LATS2; MOB proteins; kd; MST1/2; Large tumour suppressor kinase; NDR; wt
• ISSN: 0898-6568; 1873-3913
• DOI: 10.1016/j.cellsig.2016.02.012

By controlling the YAP1 proto-oncoprotein Hippo signalling plays important roles in cancer-associated processes. Current evidence suggests that the Hippo kinases MST1/2 together with the MOB1 scaffold protein promote the formation of active MOB1/LATS complexes which phosphorylate and thereby inhibit YAP1. However, the regulatory mechanisms of MST1/2-MOB1-LATS signalling are currently underinvestigated. Therefore, we studied LATS2 variants carrying specific modifications that mimic gain or loss of phosphorylation and/or abolish MOB1/LATS2 interactions. We discovered that Ser872 T-loop and Thr1041 hydrophobic motif (HM) phosphorylation of LATS2 is essential for LATS2 activation. MST1/2 phosphorylate LATS2 on Thr1041, but not Ser872, while MOB1 binding to LATS2 supports both phosphorylation events. Significantly, LATS2-PIF, a LATS2 variant containing the PRK2 HM, acts as a hyperactive LATS2 kinase that efficiently phosphorylates YAP1 and inhibits the transcriptional co-activity of YAP1. This inhibitory function of LATS2-PIF is dependent on LATS2 kinase activity, while MOB1/LATS2 and YAP1/LATS2 complex formation is dispensable, suggesting that elevated LATS2 kinase activity can be sufficient to oppose YAP1. Taken together, our characterisation of LATS2 variants uncovers novel insights into the regulation of LATS kinases in Hippo signalling.
•LATS2 activity depends on Ser872 and Thr1041 phosphorylation in the T-loop and HM.•The Hippo kinases MST1/2 phosphorylate LATS2 on Thr1041 in the HM, but not Ser872.•LATS2 activation requires binding of MOB1 to the N-terminal regulatory region.•Constitutively hyperactive LATS2-PIF functions independently of MOB1 binding.•LATS2-PIF phosphorylates and inhibits YAP1 independently of stable YAP1 binding.