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. 2018 Jun 6;8:8680. doi: 10.1038/s41598-018-27008-w

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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HIV-1 Tat disturbs the mitotic spindle localization of RPS3. (A) Mitotic defect in Lenti-Tat-transduced HeLa cells. HeLa cells were transduced with the control virus or Lenti-Tat and stained with an anti-Tat or anti-α-tubulin antibody. Localization of α-tubulin in the mitotic spindle and chromosome segregation were defective in the presence of Tat. Control, n = 60; Tat, n = 139. Experiments were repeated three times. (B) HIV-1 Tat disturbs the localization of RPS3 in the mitotic spindle. HeLa cells were transduced with the control virus or Lenti-Tat and stained with anti-Tat and anti-RPS3 antibodies. Control, n = 41; Tat, n = 73. Experiments were repeated two times. (C) Disruption of RPS3 localization in the mitotic spindle by HIV-1 Tat requires binding to RPS3 through its basic domain. HeLa cells were transfected with the plasmid expressing GST, GST-Tat, or GST-Tatm3, and the localization of RPS3 in the mitotic spindle was examined by immunostaining with antibodies against RPS3 and GST. GST-Tatm3 lacking the RPS3-binding domain did not inhibit the localization of RPS3 in the mitotic spindle, similar to GST, while wild-type Tat disturbed the localization of RPS3. GST, n = 22; GST-Tatm3, n = 25; GST-Tat, n = 32.