Cloning and tissue distribution of the ATP-binding cassette subfamily G member 2 gene in the marine pufferfish Takifugu
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ORIGINAL ARTICLE Chemistry and Biochemistry
Cloning and tissue distribution of the ATP‑binding cassette subfamily G member 2 gene in the marine pufferfish Takifugu rubripes Takuya Matsumoto1 · Saemi Kitajima2 · Chisato Yamamoto2 · Mitsuru Aoyagi2 · Yoshiharu Mitoma2 · Hiroyuki Harada2 · Yuji Nagashima3 Received: 10 April 2020 / Accepted: 10 July 2020 © Japanese Society of Fisheries Science 2020
Abstract In this study, we cloned the ATP-binding cassette (ABC) subfamily G member 2 gene (Abcg2 gene; breast cancer resistance protein) from the liver of the marine pufferfish Takifugu rubripes, and examined its tissue distribution and gene structure. The full-length complementary DNA (cDNA) was 1977 base pairs (bp), comprising 15 exons in chromosome 17, and was assigned to the teleost Abcg2b paralogue. The open reading frame was 1839 bp in length, and encodes 612 amino acids constituting Walker A, Q-loop, C motif, Walker B, D-loop, and H-loop sequences in the first half and six transmembrane domains in the second half. Gene expression of Abcg2 was predominantly found in the intestines, followed by the liver and kidneys. The Kozak motif and the polyadenylation signal sequences were at respective ends of the transcript. The putative core promoter sequences including the transcription factor II B recognition elements, the TATA box, the initiator element, and the downstream core promoter element were found in the transcript and its upstream genomic DNA sequence. The hypoxia response element and the estrogen response element were located in the upstream genomic DNA sequence, suggesting that the pufferfish Abcg2 gene possesses the same regulation mechanism of transcription as the human ABCG2 gene. Keywords Pufferfish · Takifugu rubripes · ATP-binding cassette sub-family G member 2 · Breast cancer resistance protein · Transporter · Tissue distribution · Real-time polymerase chain reaction · Splicing variant
Introduction The ATP-binding cassette (ABC) transporter family comprises membrane proteins with an ATP-binding domain that induces active transport using ATP hydrolysis (Dean et al. 2001). The ABC transporter family is divided into eight subfamilies (ABCA to ABCH), all of which actively transport
a wide variety of substrates such as sugars, amino acids, peptides, xenobiotics, lipophilic low molecular weight compounds, and physiologically active substances (Sarkadi et al. 2006). The human ABCG2 gene belongs to subfamily G; it was first discovered in a human MCF-7 breast cancer cell line that showed anticancer drug resistance, and the ABC transporter protein encoded by this gene was named the
* Takuya Matsumoto takuya62@pu‑hiroshima.ac.jp
Yuji Nagashima yuji‑[email protected]
Saemi Kitajima [email protected]
1
Department of Health Sciences, Faculty of Human Culture and Science, Prefectural University of Hiroshima, Ujina‑higashi 1‑1‑71, Minami‑ku, Hiroshima 734‑8558, Japan
2
Department of Environmental Sciences, Faculty of Life and Environmental Sciences, Prefectural University of Hiroshima, Nanatsu
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