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Identification of a Metacaspase Gene in the Bloom-Forming Dinoflagellate Prorocentrum minimum and its Putative Function Involved in Programmed Cell Death

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Abstract

Programmed cell death (PCD) in dinoflagellates has been introduced as a new concept that facilitates the demise of harmful algal blooms. Metacaspases (MCAs) play a role in PCD, but their function in dinoflagellates is unclear. Here, we cloned a novel MCA gene (PmMCA) from the harmful dinoflagellate Prorocentrum minimum and examined its molecular characteristics and gene expression during cell death. The gene was encoded in the nuclear genome with two introns. The putative protein contained 288 amino acids and three conserved MCA signature motifs. Phylogenetic analysis showed that PmMCA may have the same ancestor as other dinoflagellates. PmMCA expression and cell apoptosis were significantly induced under copper exposure, considerably affecting cell growth. These results suggest that PmMCA could be involved in PCD triggered by copper stress.

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Abbreviations

aa:

Amino acids

BLAST:

Basic local alignment search tool

CTAB:

Cetyltrimethylammonium bromide

EC50 :

Median effective concentration

EST:

Expressed sequence tag

HABs:

Harmful algal blooms

MCA:

Metacaspase

M13F:

M13 forward

M13R:

M13 reverse

NJ:

Neighbor-joining

ORF:

Open reading frame

PCD:

Programmed cell death

PI:

Propidium iodide

Pm:

Prorocentrum minimum

RACE:

Rapid amplification of cDNA ends

UTR:

Untranslated region

DinoSL:

Dinoflagellate spliced leader

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Acknowledgements

This work was supported by the National Research Foundation of Korea (NRF) Grant funded by the Korea government (MSIT) (No. 2020R1A2C2013373), and a part of the project titled ‘Development of hull adherent organism management technology (20210651),’ funded by the Ministry of Oceans and Fisheries, Korea.

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  1. Hui Wang

    Present address: Hunan Province Key Laboratory of Typical Environmental Pollution and Health Hazards, School of Public Health, University of South China, Hengyang, 421001, China

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  1. Department of Biotechnology, Sangmyung University, Seoul, 03016, South Korea

    Hui Wang & Jang-Seu Ki

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HW performed the study and drafted the manuscript; JSK revised and edited the manuscript.

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Correspondence to Jang-Seu Ki.

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Supplementary Information

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284_2021_2617_MOESM1_ESM.tif

Supplementary file1 Multiple sequence alignments (A) and homoly matrix (B) of deduced amino acid sequences of PmMCA with those of other MCAs. Amino acid sequences from 10 MCAs share similar residues. A short linker was present between P20 and P10 subunits. The signature motifs were marked with black box. GenBank accession numbers of aligned proteins are as follows: Margalefidinium polykrikoides, AVD29967; Microcystis aeruginosa, WP_002768321; Gregarina niphandrodes, XP_011130889; Fistulifera solaris, GAX24014; Coccomyxa subellipsoidea, XP_005649943; Gonium pectorale, KXZ52917; Erythranthe guttata, XP_012858139; Zostera marina, KMZ57843; Prorocentrum micans, comp12808_c0 was obtained from P. micans transcriptome data. (TIF 2500 kb)

284_2021_2617_MOESM2_ESM.tif

Supplementary file2 The maximum-likelihood tree of deduced amino acid sequence of PmMCA. The phylogenetic position of present P. minimum is marked with a red color. The amino acid sequences used in this study were obtained from GenBank database. (TIF 893 kb)

Supplementary file3 (XLSX 27 kb)

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Wang, H., Ki, JS. Identification of a Metacaspase Gene in the Bloom-Forming Dinoflagellate Prorocentrum minimum and its Putative Function Involved in Programmed Cell Death. Curr Microbiol 78, 3577–3585 (2021). https://doi.org/10.1007/s00284-021-02617-3

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