Role of the CTCF binding site in Human T-Cell Leukemia Virus-1 pathogenesis.
During HTLV-1 infection, the virus integrates into the host cell genome as a provirus with a single CCCTC binding protein (CTCF) binding site (vCTCF-BS), which acts as an insulator between transcriptionally active and inactive regions. Previous studies have shown that the vCTCF-BS is important for m...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2025-06-01
|
| Series: | PLoS Pathogens |
| Online Access: | https://doi.org/10.1371/journal.ppat.1012293 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849688324040556544 |
|---|---|
| author | Ancy Joseph Xiaogang Cheng John Harding Jacob Al-Saleem Patrick Green Malachi Griffith Deborah Veis Daniel A Rauch Lee Ratner |
| author_facet | Ancy Joseph Xiaogang Cheng John Harding Jacob Al-Saleem Patrick Green Malachi Griffith Deborah Veis Daniel A Rauch Lee Ratner |
| author_sort | Ancy Joseph |
| collection | DOAJ |
| description | During HTLV-1 infection, the virus integrates into the host cell genome as a provirus with a single CCCTC binding protein (CTCF) binding site (vCTCF-BS), which acts as an insulator between transcriptionally active and inactive regions. Previous studies have shown that the vCTCF-BS is important for maintenance of chromatin structure, regulation of viral expression, and DNA and histone methylation. Here, we show that the vCTCF-BS also regulates viral infection and pathogenesis in vivo in a humanized (Hu) mouse model of adult T-cell leukemia/lymphoma. Three cell lines were used to initiate infection of the Hu-mice, i) HTLV-1-WT which carries an intact HTLV-1 provirus genome, ii) HTLV-1-CTCF, which contains a provirus with a mutated vCTCF-BS which abolishes CTCF binding, and a stop codon immediately upstream of the mutated vCTCF-BS which deletes the last 23 amino acids of the p12 gene, and iii) HTLV-1-p12stop that contains the intact vCTCF-BS, but retains the same stop codon in p12 as in the HTLV-1-CTCF cell line. Hu-mice were infected with mitomycin-treated or irradiated HTLV-1 producing cell lines. There was a delay in pathogenicity when Hu-mice were infected with the HTLV-1-CTCF virus compared to mice infected with either HTLV-1-p12 stop or HTLV-1-WT virus. Proviral load (PVL), spleen weights, and CD4 T cell counts were significantly lower in HTLV-1-CTCF infected mice compared to HTLV-1-p12stop infected mice. Furthermore, we found a direct correlation between the PVL in peripheral blood and death of HTLV-1-CTCF infected mice. In cell lines, we found that the vCTCF-BS regulates Tax expression in a time-dependent manner. The scRNAseq analysis of splenocytes from infected mice suggests that the vCTCF-BS plays an important role in activation and expansion of T lymphocytes in vivo. Overall, these findings indicate that the vCTCF-BS regulates Tax expression, proviral load, and HTLV pathogenicity in vivo. |
| format | Article |
| id | doaj-art-af66c8fd7e3d4ff4ac0ccf8eee084d51 |
| institution | DOAJ |
| issn | 1553-7366 1553-7374 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj-art-af66c8fd7e3d4ff4ac0ccf8eee084d512025-08-20T03:22:03ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742025-06-01216e101229310.1371/journal.ppat.1012293Role of the CTCF binding site in Human T-Cell Leukemia Virus-1 pathogenesis.Ancy JosephXiaogang ChengJohn HardingJacob Al-SaleemPatrick GreenMalachi GriffithDeborah VeisDaniel A RauchLee RatnerDuring HTLV-1 infection, the virus integrates into the host cell genome as a provirus with a single CCCTC binding protein (CTCF) binding site (vCTCF-BS), which acts as an insulator between transcriptionally active and inactive regions. Previous studies have shown that the vCTCF-BS is important for maintenance of chromatin structure, regulation of viral expression, and DNA and histone methylation. Here, we show that the vCTCF-BS also regulates viral infection and pathogenesis in vivo in a humanized (Hu) mouse model of adult T-cell leukemia/lymphoma. Three cell lines were used to initiate infection of the Hu-mice, i) HTLV-1-WT which carries an intact HTLV-1 provirus genome, ii) HTLV-1-CTCF, which contains a provirus with a mutated vCTCF-BS which abolishes CTCF binding, and a stop codon immediately upstream of the mutated vCTCF-BS which deletes the last 23 amino acids of the p12 gene, and iii) HTLV-1-p12stop that contains the intact vCTCF-BS, but retains the same stop codon in p12 as in the HTLV-1-CTCF cell line. Hu-mice were infected with mitomycin-treated or irradiated HTLV-1 producing cell lines. There was a delay in pathogenicity when Hu-mice were infected with the HTLV-1-CTCF virus compared to mice infected with either HTLV-1-p12 stop or HTLV-1-WT virus. Proviral load (PVL), spleen weights, and CD4 T cell counts were significantly lower in HTLV-1-CTCF infected mice compared to HTLV-1-p12stop infected mice. Furthermore, we found a direct correlation between the PVL in peripheral blood and death of HTLV-1-CTCF infected mice. In cell lines, we found that the vCTCF-BS regulates Tax expression in a time-dependent manner. The scRNAseq analysis of splenocytes from infected mice suggests that the vCTCF-BS plays an important role in activation and expansion of T lymphocytes in vivo. Overall, these findings indicate that the vCTCF-BS regulates Tax expression, proviral load, and HTLV pathogenicity in vivo.https://doi.org/10.1371/journal.ppat.1012293 |
| spellingShingle | Ancy Joseph Xiaogang Cheng John Harding Jacob Al-Saleem Patrick Green Malachi Griffith Deborah Veis Daniel A Rauch Lee Ratner Role of the CTCF binding site in Human T-Cell Leukemia Virus-1 pathogenesis. PLoS Pathogens |
| title | Role of the CTCF binding site in Human T-Cell Leukemia Virus-1 pathogenesis. |
| title_full | Role of the CTCF binding site in Human T-Cell Leukemia Virus-1 pathogenesis. |
| title_fullStr | Role of the CTCF binding site in Human T-Cell Leukemia Virus-1 pathogenesis. |
| title_full_unstemmed | Role of the CTCF binding site in Human T-Cell Leukemia Virus-1 pathogenesis. |
| title_short | Role of the CTCF binding site in Human T-Cell Leukemia Virus-1 pathogenesis. |
| title_sort | role of the ctcf binding site in human t cell leukemia virus 1 pathogenesis |
| url | https://doi.org/10.1371/journal.ppat.1012293 |
| work_keys_str_mv | AT ancyjoseph roleofthectcfbindingsiteinhumantcellleukemiavirus1pathogenesis AT xiaogangcheng roleofthectcfbindingsiteinhumantcellleukemiavirus1pathogenesis AT johnharding roleofthectcfbindingsiteinhumantcellleukemiavirus1pathogenesis AT jacobalsaleem roleofthectcfbindingsiteinhumantcellleukemiavirus1pathogenesis AT patrickgreen roleofthectcfbindingsiteinhumantcellleukemiavirus1pathogenesis AT malachigriffith roleofthectcfbindingsiteinhumantcellleukemiavirus1pathogenesis AT deborahveis roleofthectcfbindingsiteinhumantcellleukemiavirus1pathogenesis AT danielarauch roleofthectcfbindingsiteinhumantcellleukemiavirus1pathogenesis AT leeratner roleofthectcfbindingsiteinhumantcellleukemiavirus1pathogenesis |