Distinct roles of two alternative splice variants of matrilin-2 in protein oligomerization and proteolysis

Li,Longxuan; Zhang,Liangqing; Shao,Yiming; Wang,Guirong; Gong,Rujun; Wang,Zhengke; Peng,Jinwu; Wang,Shuhua; Genochio,David; Zhao,Bin; Luo,Junming

doi:10.3892/mmr.2012.1056

Distinct roles of two alternative splice variants of matrilin-2 in protein oligomerization and proteolysis

Authors:
- Longxuan Li
- Liangqing Zhang
- Yiming Shao
- Guirong Wang
- Rujun Gong
- Zhengke Wang
- Jinwu Peng
- Shuhua Wang
- David Genochio
- Bin Zhao
- Junming Luo
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Affiliations: Department of Neurology, Guangdong Medical College Affiliated Hospital, Zhanjiang 524001, P.R. China, Department of Anesthesiology, Guangdong Medical College Affiliated Hospital, Zhanjiang 524001, P.R. China, Department of Neurology, Guangdong Medical College Affiliated Hospital, Zhanjiang 524001, P.R. China, Department of Surgery, SUNY Upstate Medical University, NY, USA, Department of Internal Medicine, Brown Medical School/Rhode Island Hospital, Providence, RI, USA, Department of Orthopaedic Research, Brown Medical School/Rhode Island Hospital, Providence, RI, USA, Department of Pathology, Xiangya Medical School, Central South University, Changsha 410073, Hunan, P.R. China, Kidney Institute, Department of Medicine, University of Kansas, Kansas City, MO, USA, VA Medical Center at Kansas City Missouri, Kansas City, MO, USA
Published online on: August 30, 2012 https://doi.org/10.3892/mmr.2012.1056
Pages: 1204-1210

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Abstract

Matrilin-2 (matn2) contains a unique domain, between the second von Willebrand factor A (vWFA) domain and the C-terminal coiled-coil domain, with no sequence homology with other family members. Complementary DNA (cDNA) sequence analysis of matn2 expression in both mice and humans revealed an alternative splice site in the region of the unique domain, which forms a short and a long splicing variant (containing an additional 19 amino acids). However, the expression heterogeneity of the alternative spliced variants, and the roles of the unique domain in oligomerization and proteolysis of matn2 are unknown. In this study, we examined the expression of the two alternative splice variants of matn2 in several skeletal and non-skeletal tissues by reverse transcription-polymerase chain reaction. Both splice variants of matn2 were detected at the mRNA level in all tissues studied. To explore the biochemical significance, several minigene constructs containing the second vWFA domain, the unique domain (with either a long or short form) and the coiled-coil domain of mouse mini matn2 were generated. Ectopic expression of these constructs demonstrated that the long form of matn2 is capable of self-assembling into several oligomeric forms, including a tetramer, trimer, pentamer or multimer; but the short form is only capable of forming a tetramer, trimer or dimer. Moreover, we observed that the splice variants of matn2 are important in modulating matn2 cleavage when co-expressed with matrilin-1 or matrilin-3. These results indicate that the two alternative splice variants have distinct roles in the processes of post-translational modification of matn2, which may have an impact on the homeostasis of the matrilin filamentous network of the extracellular matrix.

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