title: Splice variant insertions in the C-terminus impairs YAP's transactivation domain.
abstract: The yes-associated protein (YAP) is a key effector of the mammalian Hippo signaling pathway. YAP has eight known alternately spliced isoforms and these are widely expressed across multiple tissues. Variable effects have been ascribed to different YAP isoforms by inducing their expression in cells, but whether these differences are due to variability in the transcriptional potency of individual YAP isoforms has not been addressed. Indeed a systematic comparison of the transcriptional potencies of YAP isoforms has not been done. To address this, using overexpression and transcriptional reporter analyses we investigated the transcriptional activities of several human YAP isoforms and determined the effects of the splice variant insertions within the transactivation domain on its transcriptional potency. Utilising full-length coding sequence constructs we determined that the number of WW domains and disruption of the leucine zipper motif within YAP's transactivation domain both contribute to transcriptional activity. Notably, disruption of YAP's leucine zipper had a greater effect on transcriptional activity than the absence of the second WW domain. UsingGAL4-YAP transcriptional activation domain fusion proteins we found that disruption of the leucine zipper significantly decreased YAP's transcriptional activity in several cell lines. Our data indicates that expression of different YAP isoforms with varying transcriptional potencies may enable fine control of Hippo pathway signaling. Furthermore the specific isoform being utilised should be taken into consideration when interpreting published data or when designing experiments to ascribe YAP's function.
authors.affiliation: School of Chemistry and Biochemistry, University of Western Australia, WA 6009, Australia.
authors.forename: Megan L
authors.initials: ML
authors.lastname: Finch-Edmondson
authors.affiliation: School of Chemistry and Biochemistry, University of Western Australia, WA 6009, Australia; Harry Perkins Institute of Medical Research, WA 6009, Australia.
authors.forename: Robyn P
authors.initials: RP
authors.lastname: Strauss
authors.affiliation: School of Pathology and Laboratory Medicine, University of Western Australia, WA 6009, Australia.
authors.forename: Joshua S
authors.initials: JS
authors.lastname: Clayton
authors.affiliation: School of Chemistry and Biochemistry, University of Western Australia, WA 6009, Australia; Harry Perkins Institute of Medical Research, WA 6009, Australia.
authors.forename: George C
authors.initials: GC
authors.lastname: Yeoh
authors.affiliation: School of Chemistry and Biochemistry, University of Western Australia, WA 6009, Australia; School of Health Sciences, The University of Notre Dame Australia, WA 6959, Australia.
authors.forename: Bernard A
authors.initials: BA
authors.lastname: Callus
keywords: Alternate splicing; Con, control; EV, empty vector; Luciferase assay; TAD, transcriptional activation domain; Transcriptional co-activator; WT, wild-type; YAP, yes-associated protein; Yes-associated protein (YAP); aa:, Amino acid; hYAP:, human YAP; mYAP:, mouse YAP