Vitellogenin receptor transports the 30K protein LP1 without cell-penetrating peptide, into the oocytes of the silkworm, Bombyx mori
Vitellogenin receptors (VgRs) transport vitellogenin (Vg) into oocytes, thereby promoting egg growth and embryonic development. VgRs recognize and transport multiple ligands in oviparous animals, but their role in insects is rarely reported. In this study, we investigated whether Bombyx mori VgR (Bm...
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Frontiers Media S.A.
2023-01-01
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| Series: | Frontiers in Physiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphys.2023.1117505/full |
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| author | Yinying Xu Yinying Xu Guanwang Shen Guanwang Shen Guanwang Shen Guanwang Shen Jinxin Wu Jinxin Wu Jinxin Wu Xueqin Mao Xueqin Mao Linbang Jia Linbang Jia Yan Zhang Yan Zhang Yan Zhang Yan Zhang Qingyou Xia Qingyou Xia Qingyou Xia Qingyou Xia Ying Lin Ying Lin Ying Lin Ying Lin |
| author_facet | Yinying Xu Yinying Xu Guanwang Shen Guanwang Shen Guanwang Shen Guanwang Shen Jinxin Wu Jinxin Wu Jinxin Wu Xueqin Mao Xueqin Mao Linbang Jia Linbang Jia Yan Zhang Yan Zhang Yan Zhang Yan Zhang Qingyou Xia Qingyou Xia Qingyou Xia Qingyou Xia Ying Lin Ying Lin Ying Lin Ying Lin |
| author_sort | Yinying Xu |
| collection | DOAJ |
| description | Vitellogenin receptors (VgRs) transport vitellogenin (Vg) into oocytes, thereby promoting egg growth and embryonic development. VgRs recognize and transport multiple ligands in oviparous animals, but their role in insects is rarely reported. In this study, we investigated whether Bombyx mori VgR (BmVgR) binds and transports lipoprotein-1 (BmLP1) and lipoprotein-7 (BmLP7) of the 30 kDa lipoproteins (30 K proteins), which are essential for egg formation and embryonic development in B. mori. Protein sequence analysis showed BmLP7, similar to reported lipoprotein-3 (BmLP3), contains the cell-penetrating peptides and Cysteine position, while BmLP1 has not. Assays using Spodoptera frugiperda ovary cells (sf9) indicated the direct entry of BmLP7 into the cells, whereas BmLP1 failed to enter. However, co-immunoprecipitation (Co-IP) assays indicated that BmVgR could bind BmLP1. Western blotting and immunofluorescence assays further revealed that over-expressed BmVgR could transport BmLP1 into sf9 cells. Co-IP assays showed that SE11C (comprising LBD1+EGF1+OTC domains of BmVgR) or SE22C (comprising LBD2+EGF2+OTC domains of BmVgR) could bind BmLP1. Over-expressed SE11C or SE22C could also transport BmLP1 into sf9 cells. Western blotting revealed that the ability of SE11C to transport BmLP1 might be stronger than that of SE22C. In the vit mutant with BmVgR gene mutation (vit/vit), SDS-PAGE and western blotting showed the content of BmLP1 in the ovary, like BmVg, was lower than that in the normal silkworm. When transgenic with hsp70 promoter over-expressed BmVgR in the vit mutant, we found that the phenotype of the vit mutant was partly rescued after heat treatment. And contents of BmLP1 and BmVg in vit mutant over-expressed BmVgR were higher than in the vit mutant. We conclude that BmVgR and its two repeat domains could bind and transport BmLP1 into the oocytes of the silkworm, besides BmVg. These results will provide a reference for studying the molecular mechanism of VgR transporting ligands in insects. |
| first_indexed | 2024-04-10T20:16:06Z |
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| spelling | doaj.art-dd6381b2dc68492996916ab3a07d3af32023-01-26T05:38:56ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2023-01-011410.3389/fphys.2023.11175051117505Vitellogenin receptor transports the 30K protein LP1 without cell-penetrating peptide, into the oocytes of the silkworm, Bombyx moriYinying Xu0Yinying Xu1Guanwang Shen2Guanwang Shen3Guanwang Shen4Guanwang Shen5Jinxin Wu6Jinxin Wu7Jinxin Wu8Xueqin Mao9Xueqin Mao10Linbang Jia11Linbang Jia12Yan Zhang13Yan Zhang14Yan Zhang15Yan Zhang16Qingyou Xia17Qingyou Xia18Qingyou Xia19Qingyou Xia20Ying Lin21Ying Lin22Ying Lin23Ying Lin24State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, ChinaBiological Science Research Center Southwest University, Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, ChinaBiological Science Research Center Southwest University, Chongqing, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City & Southwest University, Chongqing, ChinaChongqing Key Laboratory of Sericultural Science, Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, ChinaBiological Science Research Center Southwest University, Chongqing, ChinaChongqing Key Laboratory of Sericultural Science, Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, ChinaBiological Science Research Center Southwest University, Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, ChinaBiological Science Research Center Southwest University, Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, ChinaBiological Science Research Center Southwest University, Chongqing, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City & Southwest University, Chongqing, ChinaChongqing Key Laboratory of Sericultural Science, Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, ChinaBiological Science Research Center Southwest University, Chongqing, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City & Southwest University, Chongqing, ChinaChongqing Key Laboratory of Sericultural Science, Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, ChinaBiological Science Research Center Southwest University, Chongqing, ChinaIntegrative Science Center of Germplasm Creation in Western China (Chongqing) Science City & Southwest University, Chongqing, ChinaChongqing Key Laboratory of Sericultural Science, Chongqing, ChinaVitellogenin receptors (VgRs) transport vitellogenin (Vg) into oocytes, thereby promoting egg growth and embryonic development. VgRs recognize and transport multiple ligands in oviparous animals, but their role in insects is rarely reported. In this study, we investigated whether Bombyx mori VgR (BmVgR) binds and transports lipoprotein-1 (BmLP1) and lipoprotein-7 (BmLP7) of the 30 kDa lipoproteins (30 K proteins), which are essential for egg formation and embryonic development in B. mori. Protein sequence analysis showed BmLP7, similar to reported lipoprotein-3 (BmLP3), contains the cell-penetrating peptides and Cysteine position, while BmLP1 has not. Assays using Spodoptera frugiperda ovary cells (sf9) indicated the direct entry of BmLP7 into the cells, whereas BmLP1 failed to enter. However, co-immunoprecipitation (Co-IP) assays indicated that BmVgR could bind BmLP1. Western blotting and immunofluorescence assays further revealed that over-expressed BmVgR could transport BmLP1 into sf9 cells. Co-IP assays showed that SE11C (comprising LBD1+EGF1+OTC domains of BmVgR) or SE22C (comprising LBD2+EGF2+OTC domains of BmVgR) could bind BmLP1. Over-expressed SE11C or SE22C could also transport BmLP1 into sf9 cells. Western blotting revealed that the ability of SE11C to transport BmLP1 might be stronger than that of SE22C. In the vit mutant with BmVgR gene mutation (vit/vit), SDS-PAGE and western blotting showed the content of BmLP1 in the ovary, like BmVg, was lower than that in the normal silkworm. When transgenic with hsp70 promoter over-expressed BmVgR in the vit mutant, we found that the phenotype of the vit mutant was partly rescued after heat treatment. And contents of BmLP1 and BmVg in vit mutant over-expressed BmVgR were higher than in the vit mutant. We conclude that BmVgR and its two repeat domains could bind and transport BmLP1 into the oocytes of the silkworm, besides BmVg. These results will provide a reference for studying the molecular mechanism of VgR transporting ligands in insects.https://www.frontiersin.org/articles/10.3389/fphys.2023.1117505/fullvitellogenin receptorlipoprotein 130k proteinstransportBombyx mori |
| spellingShingle | Yinying Xu Yinying Xu Guanwang Shen Guanwang Shen Guanwang Shen Guanwang Shen Jinxin Wu Jinxin Wu Jinxin Wu Xueqin Mao Xueqin Mao Linbang Jia Linbang Jia Yan Zhang Yan Zhang Yan Zhang Yan Zhang Qingyou Xia Qingyou Xia Qingyou Xia Qingyou Xia Ying Lin Ying Lin Ying Lin Ying Lin Vitellogenin receptor transports the 30K protein LP1 without cell-penetrating peptide, into the oocytes of the silkworm, Bombyx mori Frontiers in Physiology vitellogenin receptor lipoprotein 1 30k proteins transport Bombyx mori |
| title | Vitellogenin receptor transports the 30K protein LP1 without cell-penetrating peptide, into the oocytes of the silkworm, Bombyx mori |
| title_full | Vitellogenin receptor transports the 30K protein LP1 without cell-penetrating peptide, into the oocytes of the silkworm, Bombyx mori |
| title_fullStr | Vitellogenin receptor transports the 30K protein LP1 without cell-penetrating peptide, into the oocytes of the silkworm, Bombyx mori |
| title_full_unstemmed | Vitellogenin receptor transports the 30K protein LP1 without cell-penetrating peptide, into the oocytes of the silkworm, Bombyx mori |
| title_short | Vitellogenin receptor transports the 30K protein LP1 without cell-penetrating peptide, into the oocytes of the silkworm, Bombyx mori |
| title_sort | vitellogenin receptor transports the 30k protein lp1 without cell penetrating peptide into the oocytes of the silkworm bombyx mori |
| topic | vitellogenin receptor lipoprotein 1 30k proteins transport Bombyx mori |
| url | https://www.frontiersin.org/articles/10.3389/fphys.2023.1117505/full |
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