Identification of whirlin domains interacting with espin: A study of the mechanism of Usher syndrome type II

Wang,Le; Wei,Bo; Fu,Xueqi; Wang,Yuchen; Sui,Yuan; Ma,Junfeng; Gong,Xianhui; Hao,Jilong; Xing,Shu

doi:10.3892/mmr.2019.10728

Identification of whirlin domains interacting with espin: A study of the mechanism of Usher syndrome type II

Authors:
- Le Wang
- Bo Wei
- Xueqi Fu
- Yuchen Wang
- Yuan Sui
- Junfeng Ma
- Xianhui Gong
- Jilong Hao
- Shu Xing
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Affiliations: Department of Ophthalmology, First Hospital, Jilin University, Changchun, Jilin 130021, P.R. China, Department of Neurosurgery, China‑Japan Union Hospital, Jilin University, Changchun, Jilin 130033, P.R. China, Edmond H. Fischer Signal Transduction Laboratory, School of Life Sciences, Jilin University, Changchun, Jilin 130012, P.R. China, Department of Opthalmology, Eye Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, P.R. China
Published online on: October 7, 2019 https://doi.org/10.3892/mmr.2019.10728
Pages: 5111-5117
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Usher syndrome is the most common condition of combined blindness and deafness and is classified into three types (USH1‑USH3). USH2 is the most commonly diagnosed of all Usher syndrome cases. There are three identified proteins (usherin, GPR98 and whirlin) that form the USH2 complex. Defects in any of these proteins may cause failure in the formation of the USH2 complex, which is the primary cause of USH2. Whirlin is a scaffold protein and is essential for the assembly of the USH2 protein complex. It has been reported that espin is an interacting partner protein for whirlin. However, which fragment of whirlin interacts with espin remains unclear. In the present study, whirlin N‑ and C‑terminal fragments in the pEGFP‑C2 vectors were constructed. The recombinant plasmids were transfected into COS‑7 cells to observe the co‑localization by confocal laser scanning microscopy. The interactions between whirlin and espin were investigated by co‑immunoprecipitation using the 293 cell line. It was demonstated that only the whirlin N‑terminal fragment was able to interact with espin and the PR (proline‑rich) region in whirlin may be important for the interaction. However, the present study did not investigate the interaction between whirlin and espin without the PR domain which warrants future research. Our findings elucidated a primary mechanism of interaction between whirlin and espin, which are crucial for further study on the USH2 complex and USH2 pathogenesis.

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