Abstract
Southern rice black-streaked dwarf virus (SRBSDV), an insect- and plant-infecting reovirus of the genus Fijivirus, induced the formation of virus-containing tubules in infected plant and insect vector cells. Expression of the nonstructural protein P7-1 of SRBSDV in insect cells by a recombinant baculovirus resulted in the formation of tubules with dimensions and appearance similar to those found in SRBSDV-infected cells. These tubules protruded from the cell surface, supporting the hypothesis that the P7-1 protein contains two putative transmembrane domains that are necessary for the formation of tubules. Furthermore, the self-interaction of SRBSDV P7-1 protein indicates that this protein has the capacity to form homodimers or oligomers to assemble the proposed helical symmetry structure of tubules. Taken together, our results indicate that SRBSDV P7-1 has the intrinsic ability of self-interaction to form tubules growing from the cell surface in the absence of other viral proteins.
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Acknowledgments
This work was supported by grants from the programs for the National Basic Research Program of China (No. 2010CB126203), Specialized Research Fund for the Ministry of Agriculture (No. 201003031), New Century Excellent Talents in University (No. NCET-09-0011), the National Natural Science Foundation of China (No. 31070130) and the Natural Science Foundation of Fujian Province (No. 2010J01075).
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Y. Liu and D. Jia contributed equally to this work.
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Fig. S1
Schematic diagram of the use of fusion PCR to synthesize the fragments for creating mutant P7-1 in which the TM1 or TM2 segment was deleted. (A) Segment 1-107 and segment 127-358 were amplified with primers P1 and P2, and primers P3 and P4, respectively. P2 and P3 are complementary PCR primers consisting of two fused sections (shown in red and blue) that match regions flanking the deleted domain. The two fragments of PCR products were mixed and fused together by PCR using primers P1 and P4, creating mutant P7-1, in which the TM1 was deleted. (B) Segment 1-285 and segment 304-358 were amplified with primers P1 and P5, and primers P6 and P4, respectively. P5 and P6 are complementary PCR primers consisting of two fused sections (shown in red and blue) that match regions flanking the deleted domain. The two fragments of the PCR products were mixed and fused together by PCR using primers P1 and P4, creating mutant P7-1, in which TM2 was deleted. (TIFF 134 kb)
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Liu, Y., Jia, D., Chen, H. et al. The P7-1 protein of southern rice black-streaked dwarf virus, a fijivirus, induces the formation of tubular structures in insect cells. Arch Virol 156, 1729–1736 (2011). https://doi.org/10.1007/s00705-011-1041-9
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DOI: https://doi.org/10.1007/s00705-011-1041-9