Cloning and functional analysis of c/ebpα as negative regulator of dmrt1 in Chinese tongue sole (Cynoglossus semilaevis)

Qian Wang; Rui Wang; Bo Feng; Shuo Li; Shahid Mahboob; Changwei Shao

doi:10.1016/j.gene.2020.145321

Cloning and functional analysis of c/ebpα as negative regulator of dmrt1 in Chinese tongue sole (Cynoglossus semilaevis)

Gene. 2021 Feb 5:768:145321. doi: 10.1016/j.gene.2020.145321. Epub 2020 Nov 19.

Authors

Qian Wang¹, Rui Wang¹, Bo Feng², Shuo Li³, Shahid Mahboob⁴, Changwei Shao⁵

Affiliations

¹ Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China.
² Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China.
³ Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Ningbo University, Ningbo 315211, China.
⁴ Department of Zoology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia.
⁵ Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China. Electronic address: shaocw@ysfri.ac.cn.

PMID: 33221538
DOI: 10.1016/j.gene.2020.145321

Abstract

c/ebpα is a member of the C/EBP family of transcription factors, which are involved in cell growth and differentiation and have a conserved basic leucine zipper (bZIP) domain. However, little is known about its function in sex determination and differentiation. In the present study, c/ebpα was cloned from the gonads of Chinese tongue sole (Cynoglossus semilaevis). The full-length cDNA of c/ebpα was 1583 bp, with a 198-bp 5' UTR, a 446-bp 3' UTR, and a 939-bp open reading frame encoding a 312-amino acid peptide. qRT-PCR revealed that c/ebpα was predominantly expressed in undifferentiated gonads of male C. semilaevis at 30 dpf and 60 dpf and peaked at 60 dpf. Expression levels of c/ebpα in the testis were constantly higher than those in ovaries at all developmental stages. Moreover, a dual-luciferase assay revealed that c/ebpα could negatively regulate the male-determining gene dmrt1 in vitro. These results provide fundamental information indicating that C. semilaevis c/ebpa might be involved in early gonadal differentiation and functions as a negative regulator of dmrt1 by repressing its transcription.

Keywords: Cynoglossus semilaevis; Gonadal differentiation; Testis; Transcription factor; c/ebpα; dmrt1.

MeSH terms

Animals
Base Sequence
CCAAT-Enhancer-Binding Protein-alpha / genetics*
CCAAT-Enhancer-Binding Protein-alpha / metabolism*
Cloning, Molecular / methods*
Female
Fish Proteins / genetics
Fish Proteins / metabolism*
Flatfishes / genetics
Flatfishes / growth & development*
Flatfishes / metabolism
Gene Expression Profiling
Gene Expression Regulation, Developmental
Male
Open Reading Frames
Ovary / growth & development
Ovary / metabolism
Phylogeny
Sequence Analysis, DNA / methods
Sex Determination Processes
Testis / growth & development
Testis / metabolism
Transcription Factors / metabolism*

Substances

CCAAT-Enhancer-Binding Protein-alpha
DMRT1 protein
Fish Proteins
Transcription Factors