Biochemical and molecular characterization of the SBiP1 chaperone from Symbiodinium microadriaticum CassKB8 and light parameters that modulate its phosphorylation.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science

Molecular Chaperones*/genetics ; Molecular Chaperones*/metabolism ; HSP70 Heat-Shock Proteins*/metabolism; Humans ; Phosphorylation ; Protein Binding ; Endoplasmic Reticulum Chaperone BiP ; Nucleotides/metabolism

The coding and promoter region sequences from the BiP-like protein SBiP1 from Symbiodinium microadriaticum CassKB8 were obtained by PCR, sequenced and compared with annotated sequences. The nucleotides corresponding to the full sequence were correctly annotated and the main SBiP1 features determined at the nucleotide and amino acid level. The translated protein was organized into the typical domains of the BiP/HSP70 family including a signal peptide, a substrate- and a nucleotide-binding domain, and an ER localization sequence. Conserved motifs included a highly conserved Thr513 phosphorylation site and two ADP-ribosylation sites from eukaryotic BiP's. Molecular modeling showed the corresponding domain regions and main exposed post-translational target sites in its three-dimensional structure, which also closely matched Homo sapiens BiP further indicating that it indeed corresponds to a BiP/HSP70 family protein. The gene promoter region showed at least eight light regulation-related sequences consistent with the molecule being highly phosphorylated in Thr under dark conditions and dephosphorylated upon light stimuli. We tested light parameter variations that could modulate the light mediated phosphorylation effect and found that SBiP1 Thr dephosphorylation was only significantly detected after 15-30 min light stimulation. Such light-induced dephosphorylation was observed even when dichlorophenyl dimethyl urea, a photosynthesis inhibitor, was also present in the cells during the light stimulation. Dephosphorylation occurred indistinctly under red, yellow, blue or the full visible light spectra. In additon, it was observed at a light intensity of as low as 1 μmole photon m-2 s-1. Our results indicate that: a) SBiP1 is a chaperone belonging to the BiP/HSP70 family proteins; b) its light-modulated phosphorylation/dephosphorylation most likely functions as an activity switch for the chaperone; c) this light-induced modulation occurs relatively slow but is highly sensitive to the full spectrum of visible light; and d) the light induced Thr dephosphorylation is independent of photosynthetic activity in these cells.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2023 Castillo-Medina et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

PeerJ. 2019 Aug 12;7:e7406. (PMID: 31423357)
Plant Cell Physiol. 1997 Apr;38(4):404-12. (PMID: 9177027)
Front Plant Sci. 2017 Feb 28;8:271. (PMID: 28293249)
Acta Biochim Pol. 2022 Feb 11;69(1):155-164. (PMID: 35148474)
J Exp Bot. 2005 Feb;56(412):557-65. (PMID: 15569706)
Plant Signal Behav. 2017 Mar 4;12(3):e1290039. (PMID: 28277974)
J Biol Chem. 2020 Jul 31;295(31):10689-10708. (PMID: 32518165)
Nucleic Acids Res. 2010 Jan;38(Database issue):D161-6. (PMID: 19858104)
BMC Plant Biol. 2014 Oct 01;14:260. (PMID: 25273817)
Can J Microbiol. 2014 Nov;60(11):767-75. (PMID: 25372347)
Sci Rep. 2017 Nov 3;7(1):15021. (PMID: 29101370)
Science. 2002 Dec 6;298(5600):1912-34. (PMID: 12471243)
Plant Physiol. 2011 Oct;157(2):730-41. (PMID: 21825107)
Sci Rep. 2020 Jul 9;10(1):11327. (PMID: 32647371)
Philos Trans R Soc Lond B Biol Sci. 2012 Sep 19;367(1602):2619-39. (PMID: 22889912)
J Biomol Struct Dyn. 2014;32(11):1766-79. (PMID: 24028577)
Nature. 1970 Aug 15;227(5259):680-5. (PMID: 5432063)
Microorganisms. 2021 Apr 09;9(4):. (PMID: 33918967)
J Eukaryot Microbiol. 2019 Mar;66(2):254-266. (PMID: 30027647)
Front Plant Sci. 2016 Aug 03;7:1157. (PMID: 27536312)
Nat Methods. 2022 Jun;19(6):679-682. (PMID: 35637307)
Anal Biochem. 2008 Feb 15;373(2):377-9. (PMID: 17850757)
Mol Cell Biol. 1988 Oct;8(10):4250-6. (PMID: 3141786)
Bioinformatics. 2018 Aug 1;34(15):2566-2574. (PMID: 29554239)
Nucleic Acids Res. 2021 Jan 8;49(D1):D344-D354. (PMID: 33156333)
Proteomics. 2004 Jun;4(6):1633-49. (PMID: 15174133)
Mt Sinai J Med. 2004 Oct;71(5):289-97. (PMID: 15543429)
Methods Mol Biol. 2007;365:9-22. (PMID: 17200550)
Nucleic Acids Res. 2013 Jan;41(Database issue):D344-7. (PMID: 23161676)
Nature. 2021 Aug;596(7873):583-589. (PMID: 34265844)
Sci Rep. 2016 Dec 22;6:39734. (PMID: 28004835)
Nucleic Acids Res. 2022 Jan 7;50(D1):D439-D444. (PMID: 34791371)
Semin Cell Dev Biol. 2010 Jul;21(5):472-8. (PMID: 20026282)
Infect Immun. 1994 Jun;62(6):2499-507. (PMID: 8188375)
Nat Biotechnol. 2019 Apr;37(4):420-423. (PMID: 30778233)
Cell Commun Signal. 2011 Oct 06;9:22. (PMID: 21978545)
Elife. 2016 Jul 11;5:. (PMID: 27400267)
Proc Natl Acad Sci U S A. 2014 Mar 11;111(10):3871-6. (PMID: 24567382)
BMC Biol. 2021 Apr 13;19(1):73. (PMID: 33849527)
Front Plant Sci. 2015 Dec 08;6:1090. (PMID: 26697044)
Stroke. 2019 Jan;50(1):162-171. (PMID: 30580718)
Trends Biochem Sci. 1990 Dec;15(12):483-6. (PMID: 2077689)
Commun Biol. 2018 Jul 17;1:95. (PMID: 30271976)
J Cell Biol. 2012 Aug 6;198(3):371-85. (PMID: 22869598)
Nucleic Acids Res. 1999 Jan 1;27(1):297-300. (PMID: 9847208)
J Mol Biol. 1990 Oct 5;215(3):403-10. (PMID: 2231712)
Protein Sci. 2018 Jan;27(1):14-25. (PMID: 28710774)

0 (Molecular Chaperones)
0 (HSP70 Heat-Shock Proteins)
0 (Endoplasmic Reticulum Chaperone BiP)
0 (Nucleotides)

Date Created: 20231020 Date Completed: 20231101 Latest Revision: 20231101

20240829

PMC10588850

10.1371/journal.pone.0293299

37862348