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Antwi-Adjei et al. Vessel Plus 2021;5:35 Vessel Plus
DOI: 10.20517/2574-1209.2021.48

Original Article Open Access

Furry is a component of the CCM3-GCKIII signaling

pathway

2
1
Emmanuel Antwi-Adjei , Alondra S. Burguete , Amin S. Ghabrial 1
1
Department of Pathology and Cell Biology, Columbia University Irving Medical Center, New York, NY 10032, USA.
2
Department of Neurology Columbia University Irving Medical Center, New York, NY 10032, USA.
Correspondence to: Prof. Amin S. Ghabrial, Department of Pathology and Cell biology, Columbia University Irving Medical
Center, 630 W. 168th Street, VP&S 14-401L, New York, NY 10032, USA. E-mail: asg2236@cumc.columbia.edu
How to cite this article: Antwi-Adjei E, Burguete AS, Ghabrial AS. Furry is a component of the CCM3-GCKIII signaling pathway.
Vessel Plus 2021;5:35. https://dx.doi.org/10.20517/2574-1209.2021.48

Received: 23 Mar 2021 First Decision: 15 Apr 2021 Revised: 30 Apr 2021 Accepted: 26 May 2021 First online: 26 May 2021

Academic Editor: Jun Zhang Copy Editor: Yue-Yue Zhang Production Editor: Yue-Yue Zhang

Abstract
Aim: Mutations in 3 genes encoding proteins of the Cerebral Cavernous Malformations (CCM) ternary complex
cause autosomal dominant cerebral vascular disease. Targets of CCM complex regulation have been identified;
however, the molecular mechanisms connecting CCM3 to these downstream effectors remain elusive. We aim to
determine the mechanism of CCM3 action by using a Drosophila model to elucidate the signaling pathway
downstream of CCM3. Previously, we showed that CCM3 and its binding partner, Germinal Center Kinase 3, are
required in tracheal terminal cells to prevent tube morphogenesis defects. Further, we established that GCKIII
phosphorylates and directly activates a downstream kinase, Tricornered (Drosophila STK38/38L ortholog). Here
we aim to test whether Tricornered-associated scaffolding protein, Furry, is required for CCM3-GCKIII signaling.

Methods: We utilized the FRT-FLP system to generate genetic mosaic Drosophila larvae and adults. Mitotic
recombination was induced in embryos (trachea) or larvae (wing disc). The animals were heterozygous for the
gene of interest (ccm3 or furry), but after recombination, homozygous mutant daughter cells were produced. In
addition, the GAL4-UAS system was used to express dominant negative GCKIII in wing disc cells. Mutant cells
were analyzed by brightfield and/or fluorescent microscopy.

Results: We find that wing cells mutant for ccm3, or expressing dominant negative GCKIII, produce wing hair
defects characteristic of mutations in tricornered and furry. Likewise, tracheal terminal cells mutant for furry produce
tube dilation defects characteristic of cells mutant for ccm3 or GCKIII.

© The Author(s) 2021. Open Access This article is licensed under a Creative Commons Attribution 4.0
International License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, sharing,
adaptation, distribution and reproduction in any medium or format, for any purpose, even commercially, as
long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and
indicate if changes were made.

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