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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2021.11
- 수록면
- 1,269 - 1,276 (8page)
- DOI
- 10.1007/s13258-021-01148-2
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초록· 키워드
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Background Botrytis-induced Kinase 1 (BIK1) is a receptor-like cytoplasmic kinase (RLCK) involved in the defense, growth, and development of higher plants. It interacts with various receptor-like kinases (RLKs) such as Brassinosteroid Insensitive 1 (BRI1), Flagellin Sensitive 2 (FLS2), and Perception of the Arabidopsis Danger Signal Peptide 1 (PEPR1), but little is known about signaling downstream of BIK1.
Objective In this study, we aimed to identify Arabidopsis thaliana BIK1 (AtBIK1) and Brassica rapa BIK1 (BrBIK1) interacting proteins, which is downstream signaling components in Arabidopsis. In addition, the efect of BIK1 phosphorylation on their interaction were examined.
Methods For yeast two hybrid (Y2H) screening, a B. rapa cDNA activation domain (AD) library and an A. thaliana cDNA library were used. Reverse reaction (LR) recombinations of appropriate open reading frames (AtBIK1, BrBIK1, AtRGP2, AtPATL2, AtPP7) in either pDONR207 or pDONR/zeo were performed with the split-YFP destination vectors pDESTGWVYNE and pDEST-GWVYCE to generate N- or C-terminal fusions with the N- and C-terminal yellow fuorescent protein (YFP) moieties, respectively. Recombined vectors were transformed into Agrobacterium strain GV3101. The described GSTAtBIK1, Flag-AtBIK1, and Flag-BrBIK1 constructs were used as templates for site-directed mutagenesis with a QuikChange XL Site-Directed Mutagenesis Kit (Stratagene).
Results In results, A. thaliana BIK1 (AtBIK1) displays strong autophosphorylation kinase activity on tyrosine and threonine residues, whereas B. rapa BIK1 (BrBIK1) does not exhibit autophosphorylation kinase activity in vitro. Herein, we demonstrated that four proteins (RGP2, PATL2, PP7, and SULTR4.1) interact with BrBIK1 but not AtBIK1 in a Y2H system.
To confrm interactions between BIK1 and protein candidates in Nicotiana benthamiana, BiFC analysis was performed and it was found that only BrBIK1 bound the three proteins tested. Three phosphosites, T90, T362, and T368, based on amino acid sequence alignment between AtBIK1 and BrBIK1, and performed site-directed mutagenesis (SDM) on AtBIK1 and BrBIK. S90T, P362T, and A369T mutations in BrBIK1 restored autophosphorylation kinase activity on threonine residues comparable to AtBIK1. However, T90A, T362P, and T368A mutations in AtBIK1 did not alter autophosphorylation kinase activity on threonine residues compared with wild-type AtBIK1. BiFC results showed that BIK1 mutations restored kinase activity led to the loss of the binding activity to RGP2, PATL2, or PP7 proteins.
Conclusion Phospho-BIK1 might be involved in plant innate immunity, while non-phospho BIK1 may regulate plant growth and development through interactions with RGP2, PATL2, and PP7.
목차
등록된 정보가 없습니다.
참고문헌 (21)
참고문헌 신청
Chen S / 2019 / BIK1 and ERECTA play opposing roles in both leaf and inforescence development in Arabidopsis / Front Plant Sci 10(1480):1~12
Drakakaki G / 2006 / Arabidopsis reversibly glycosylated polypeptides1 and 2are essential for pollen development / Plant Physiol 142:1480~1492
Gehl C / 2009 / New GATEWAY vectors for high throughput analyses of protein-protein interactions by bimolecular fuorescence complementation / Mol Plant 2:1051~1058
Kumanr D / 2016 / Arabidopsis thaliana RECEPTOR DEAD KINASE1functions as a positive regulator in plant responses to ABA / Mol Plant 10:223~243
Laluk K / 2011 / Biochemical and genetic requirements for function of the immune response regulator BOTRYTIS-INDUCED KINASE1 in plant growth, ethylene signaling, and PAMP-triggered immunity in Arabidopsis / Plant Cell 23:2831~2849
Drakakaki G / 2006 / Arabidopsis reversibly glycosylated polypeptides1 and 2are essential for pollen development / Plant Physiol 142:1480~1492
Gehl C / 2009 / New GATEWAY vectors for high throughput analyses of protein-protein interactions by bimolecular fuorescence complementation / Mol Plant 2:1051~1058
Kumanr D / 2016 / Arabidopsis thaliana RECEPTOR DEAD KINASE1functions as a positive regulator in plant responses to ABA / Mol Plant 10:223~243
Laluk K / 2011 / Biochemical and genetic requirements for function of the immune response regulator BOTRYTIS-INDUCED KINASE1 in plant growth, ethylene signaling, and PAMP-triggered immunity in Arabidopsis / Plant Cell 23:2831~2849
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