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Journal of Applied Phycology

, Volume 31, Issue 5, pp 2969–2979 | Cite as

Cloning and comparative studies of proliferating cell nuclear antigen (PCNA) genes for nine dinoflagellates

  • Yunyan Deng
  • Zhangxi Hu
  • Zhaoyang Chai
  • Ying Zhong TangEmail author
Article

Abstract

Proliferating cell nuclear antigen (PCNA), a co-factor of DNA polymerases δ and ε, is associated with active cell proliferation, particularly with the S phase of the eukaryotic cell cycle, and the expression of this gene has been therefore proposed as a molecular marker for estimating phytoplankton growth rate and even population dynamics. In this study, the full-length cDNA sequences of PCNA for nine dinoflagellate species (ten strains) were obtained, analyzed, and further characterized with the expression in response to alterations of growth stage and life cycle. All the ten genes have a 777 or 780 bp ORF and encode a protein of 258 or 259 amino acids, similar to PCNAs from vertebrates and plants. The deduced amino acid sequences include conservative motif characteristic to the PCNA gene family, implying conserved functions of PCNA among organisms of different taxa. Alignment analysis in combination with other sequences available in NCBI database exhibited high amino acid similarities (79.2–96.9%) among PCNAs from 17 dinoflagellate species. Phylogenetic tree derived from 37 PCNA sequences of dinoflagellates was consistent with the taxonomic affiliation of these taxa. Then, we used the cosmopolitan, toxic, and resting cyst-producing dinoflagellate Scrippsiella trochoidea as a representative of HAB-forming dinoflagellates to investigate PCNA (StPCNA) expression at different stages of growth and life cycle and the results of real-time quantitative PCR revealed clearly a stage-dependent pattern in the transcription of StPCNA. The transcripts dramatically reduced from an exponential to a stationary growth stage and then further significantly decreased from a stationary stage to dormant stage (resting cysts), a pattern in accordance with that previously reported from the dinoflagellates Alexandrium catenella and Prorocentrum donghaiense. In terms of its minimal implication, this work enhances the previous perception about the function and possible application of PCNA gene, and more importantly, our results indicated that PCNA is not silent during the dormant stage of dinoflagellates.

Keywords

Dinoflagellate Harmful algal blooms (HABs) Proliferating cell nuclear antigen (PCNA) Resting cyst Scrippsiella trochoidea 

Notes

Funding information

We acknowledge financial support from the National Science Foundation of China (grant number 41606126), the NSFC-Shandong Joint Fund for Marine Ecology and Environmental Sciences (grant number U1606404), and the National Science Foundation of China (grant numbers 41476142, 41506143, 61533011, and U1301235).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10811_2019_1809_MOESM1_ESM.docx (26 kb)
Table S1 (DOCX 26 kb)
10811_2019_1809_MOESM2_ESM.docx (26 kb)
Table S2 (DOCX 25 kb)
10811_2019_1809_MOESM3_ESM.ods (21 kb)
Table S3 (ODS 21 kb)
10811_2019_1809_MOESM4_ESM.ods (21 kb)
Table S4 (ODS 21 kb)
10811_2019_1809_Fig3_ESM.png (1.2 mb)
Fig. S1

Alignment and comparison of 18 PCNAs deduced amino acid sequences from dinoflagellates. Sequences are numbered on the right. Identical amino acid residues are black shaded, similar residues are light gray shaded. The binding sites of replication factor C (-YLAPK-) and that of DNA polymerase δ and p21 (-DSSHV-) are highlighted by arrows above alignment. One highly conserved domain with the amino acids sequence of ADFDLKLMQ is marked with triangles. Deletions are indicated by dashes. Abbreviations are the same as in Table 2 and Table S2. (PNG 1212 kb)

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High resolution image (TIF 6707 kb)
10811_2019_1809_Fig4_ESM.png (50 kb)
Fig. S2

Light microscopic observations for vegetative cell (A) and resting cyst (B) of Scrippsiella trochoidea. A. a typical swimming (with two flagella) vegetative cell with conical epitheca, round hypotheca, and apical process. Scale bar = 20 μm. B. egg or oval shaped resting cyst with a red accumulation body (arrow) and numerous surface spines. Scale bar = 20 μm. (PNG 49 kb)

10811_2019_1809_MOESM6_ESM.tif (109 kb)
High resolution image (TIF 109 kb)
10811_2019_1809_MOESM7_ESM.docx (67 kb)
Fig. S3 Standard curves in the qPCR detection showing efficiency (E) and R square values. (DOCX 67 kb)
10811_2019_1809_Fig5_ESM.png (93 kb)
Fig. S4

Phylogenetic tree of dinoflagellate species inferred from PCNA amino acid sequences based on maximum likelihood (ML) method. Number at the node represent ML bootstrap. Only bootstrap values >50 are shown. The 10 new sequences are highlighted in gray background. (PNG 92 kb)

10811_2019_1809_MOESM8_ESM.tif (334 kb)
High resolution image (TIF 334 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.CAS Key Laboratory of Marine Ecology and Environmental Sciences, Institute of OceanologyChinese Academy of SciencesQingdaoChina
  2. 2.Laboratory for Marine Ecology and Environmental ScienceQingdao National Laboratory for Marine Science and TechnologyQingdaoChina
  3. 3.Center for Ocean Mega-ScienceChinese Academy of SciencesQingdaoChina

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