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Expression characteristics of two ubiquitin/ribosomal fusion protein genes in the developing testis, accessory gonad and ovary of Chinese mitten crab, Eriocheir sinensis

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Abstract

The ubiquitin/proteasome pathway plays an important role in gametogenesis. Ubiquitin/ribosomal fusion protein is the precursor of mono-ubiquitin. We identified two ubiquitin/ribosomal fusion proteins genes in testis of Eriocheir sinensis. The ubiquitin-ribosomal proteins S27 and L40 are denoted as UBS27 and UBL40, respectively. The full length Es-UBS27 cDNA (565 bp) was cloned using PCR, based upon an initial expressed sequence tag (EST) isolated from a testicular cDNA library and the result of 3′ RACE. The sequence contained a 465-nucleotide open reading frame (ORF) that encoded a protein of 154 amino acids. It contained a Ubiquitin (UB) domain and the ribosomal protein S27. The full-length Es-UBL40 cDNA (521 bp) contained a 390-nucleotide ORF that encoded a protein of 129 amino acids. It contained a UB domain and the ribosomal protein L40. By real-time PCR, the expression level of these two UB genes is shown to change significantly in the developing testis and ovary, reaching peak expression in August and October, respectively. During accessory gonad seasonal development, the expression of UBS27 and UBL40 varied sharply and reached peak expression in November. In situ hybridization of testis and ovary, indicated that UB transcription took place widely. Our study demonstrates, for the first time, that UBS27 and UBL40 play key roles in gametogenesis and reproductive success in Eriocheir sinensis.

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Acknowledgments

This research was supported by the National Natural Science Foundation of China (No. 30972241).

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  1. School of Life Science, East China Normal University, North Zhong-Shan Road, Shanghai, China

    Qun Wang, Lili Chen, Ying Wang, Weiwei Li, Lin He & Hui Jiang

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  1. Qun Wang
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Correspondence to Qun Wang.

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Qun Wang, Lili Chen, and Ying Wang contributed equally to this work.

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11033_2012_1474_MOESM1_ESM.jpg

Fig. S1. Nucleotide and deduced amino acid sequences of Es-UBS27. The nucleotide is numbered from the first base at the 5′ end. The first methionine (M) is numbered as the first deduced amino acid. The UB sequence is shown in italics. The black circle indicates the cysteines, which comprise a zinc finger. The black oval indicates the polyadenylation sequence. The diglycine motif is bold. (JPEG 1416 kb)

11033_2012_1474_MOESM2_ESM.jpg

Fig. S2. Nucleotide and deduced amino acid sequences of Es-UBL40. The nucleotide is numbered from the first base at the 5′ end. The first methionine (M) is numbered as the first deduced amino acid. The UB sequence is shown in italics. The black circle shows the cysteines, which comprise a zinc finger. The black oval indicates the nuclear localization signal. Underlying shows the polyadenylation sequence. (JPEG 1299 kb)

11033_2012_1474_MOESM3_ESM.jpg

Fig. S3. Multiple alignment of the Es-UBS27 amino acid sequences with UBS27 from BLASTp homology search results.. Identical (*) and similar (. or :) residues are indicated. Gaps (-) were introduced to maximize the alignment. E. sinensis is marked by the black frame. (JPEG 1806 kb)

11033_2012_1474_MOESM4_ESM.jpg

Fig. S4. Multiple alignment of the Es-UBL40 amino acid sequences with UBL40 from BLASTp homology search results. Identical (*) and similar (. or :) residues are indicated. Gaps (-) were introduced to maximize the alignment. E. sinensis is marked by the black frame. (JPEG 1832 kb)

11033_2012_1474_MOESM5_ESM.jpg

Fig. S5. Neighbor-joining phylogenetic tree for UBS27 mRNA sequences reported in representative taxa. Protein abbreviations and corresponding GenBank accession numbers are as follows: Eriocheir sinensis (HM177456), Danio rerio (BC049478), Gallus gallus (NP990284), Homo sapiens (BC053371), Scylla paramamosain (GQ903729), Portunus pelagicus (EF110526), Ixodes scapularis (DQ066288), Ornithodoros parkeri (EF633956), Rana catesbeiana (BT081468), Callithrix jacchus (XM002747206), Salmo salar (BT043890), Rattus norvegicus(XM574121.3), Dermacentor variabilis (ACF35544), Mus musculus (NM024277.2), Epinephelus coioides(AF502248), Oncorhynchus mykiss(BT074005), Anoplopoma fimbria (BT083357), Rhipicephalus sanguineus (EZ406189), Bos Taurus (XM001787994.2), Pongo abelii (XM002812024), Macaca mulatta (XM001087600.2). (JPEG 345 kb)

11033_2012_1474_MOESM6_ESM.jpg

Fig. S6. Neighbor-joining phylogenetic tree for UBL40 mRNA sequences reported in representative taxa. Protein abbreviations and corresponding GenBank accession numbers are as follows: Eriocheir sinensis (HM177457), Branchiostoma belcheri (AF395864), Biomphalaria glabrata (AF268491), Crassostrea hongkongensis (GU144019), Salmo salar (NM001139743), Phlebotomus papatasi (EU032350), Homo sapiens (AF348700), Oncorhynchus mykiss (NM001124194), Danio rerio (NM001037113), Bubalus bubalis (AY842446), Ictalurus punctatus (AF401597), Gallus gallus (NM205075), Macaca mulatta (XM001118431), Rattus norvegicus (BC061544), Bufo gargarizans (DQ855628). (JPEG 271 kb)

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Wang, Q., Chen, L., Wang, Y. et al. Expression characteristics of two ubiquitin/ribosomal fusion protein genes in the developing testis, accessory gonad and ovary of Chinese mitten crab, Eriocheir sinensis . Mol Biol Rep 39, 6683–6692 (2012). https://doi.org/10.1007/s11033-012-1474-6

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