A novel multiplex RT-PCR system detects human endogenous retrovirus-K in breast cancer

Ejthadi, H. Davari; Martin, Noreen; Junying, J.; Roden, Denise A.; Lahiri, Mayurika; Warren, Philip; Murray, Paul G.; Nelson, Paul N.

doi:10.1007/s00705-004-0378-8

Cited by 23 publications

(23 citation statements)

References 25 publications

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“…As the Bogota probe packaged into HK2 particles proved to be a useful indicator of HK2 transmission, we used it next to assess whether or not cell lines other than NCCIT produce transmissible HK2 VLPs. We tested the capacity of the transmission of putative HK2 VLPs produced by the breast cancer cell line MCF-7 and the peripheral blood lymphocytes (PBLs) from patients with lymphoma, in which some recombinant HK2 viral genomes are detected, suggesting the presence of transmissible HK2 viruses (25,84). Compared to the standard particle-producing NCCIT cells, transfection of HK2 Bogota into MCF-7 and PBLs from patients with lymphoma results in the production of a small number of transmissible HK2 particles that conferred G418 resistance onto target 293T cells (Fig.…”

Section: Resultsmentioning

confidence: 99%

Human Endogenous Retrovirus Type K (HERV-K) Particles Package and Transmit HERV-K–Related Sequences

Contreras-Galindo

Kaplan

Dube

et al. 2015

J Virol

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Human endogenous retroviruses (HERV) make up 8% of the human genome. While the youngest of these retroviruses, HERV-K(HML-2), termed HK2, is able to code for all viral proteins and produce virus-like particles, it is not known if these virus particles package and transmit HK2-related sequences. Here, we analyzed the capacity of HK2 for packaging and transmitting HK2 sequences. We created an HK2 probe, termed Bogota, which can be packaged into HK2 viruses, and transfected it into cells that make HK2 particles. Supernatants of the transfected cells, which contained HK2 viral particles, then were added to target cells, and the transmissibility of the HK2 Bogota reporter was tracked by G418 resistance. Our studies revealed that contemporary HK2 virions produced by some teratocarcinoma and breast cancer cell lines, as well as by peripheral blood lymphocytes from lymphoma patients, can package HK2 Bogota probes, and these viruses transmitted these probes to other cells. After transmission, HK2 Bogota transcripts undergo reverse transcription, a step impaired by antiretroviral agents or by introduction of mutations into the probe sequences required for reverse transcription. HK2 viruses were more efficiently transmitted in the presence of HK2 Rec or HIV-1 Tat and Vif. Transmitted Bogota probes formed episomes but did not integrate into the cellular genome. Resistance to integration might explain the relatively low number of HK2 insertions that were acquired during the last 25 million years of evolution. Whether transient transmission of modern HK2 sequences, which encode two putative oncoproteins, can lead to disease remains to be studied. IMPORTANCERetroviruses invaded the genome of human ancestors over the course of millions of years, yet these viruses generally have been inactivated during evolution, with only remnants of these infectious sequences remaining in the human genome. One of these viruses, termed HK2, still is capable of producing virus particles, although these particles have been regarded as being noninfectious. Using a genetic probe derived from HK2, we have discovered that HK2 viruses produced in modern humans can package HK2 sequences and transmit them to various other cells. Furthermore, the genetic sequences packaged in HK2 undergo reverse transcription. The transmitted probe circularized in the cell and failed to integrate into the cellular genome. These findings suggest that modern HK2 viruses can package viral RNA and transmit it to other cells. Contrary to previous views, we provide evidence of an extracellular viral phase of modern HK2 viruses. We have no evidence of sustained, spreading infection.A ctively replicating retroviruses infected the germ line multiple times over the millions of years of human evolution. These sequences were transmitted vertically in a Mendelian fashion to the next generation; therefore, they became a stable part of the inherited genetic material. Relics of these retroviral infections presently make up approximately 8% of the modern human genome. These retro...

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Section: Resultsmentioning

confidence: 99%

Human Endogenous Retrovirus Type K (HERV-K) Particles Package and Transmit HERV-K–Related Sequences

Contreras-Galindo

Kaplan

Dube

et al. 2015

J Virol

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“…All samples were tested in triplicate and the mean value obtained. DNA sequencing 5 of amplicons confirmed 99% homology with ERV-3.…”

Section: To the Editormentioning

confidence: 76%

“…Peripheral blood mononuclear cells (PBMC) from 20 RA patients who fulfilled American Rheumatism Association criteria were used to extract cDNA following established protocols 5 . The average age was 61 years (range 34-85 yrs).…”

Section: To the Editormentioning

confidence: 99%

“…Seventeen patients with OA (mean age 67 yrs, range 40-92 yrs) and 27 healthy individuals (mean age 47 yrs, range 21-80 yrs) served as controls, with permission of the local ethics committee. Samples were tested in a multiplex system 5 , and optimized for the envelope region of ERV-3. In brief, ERV-3 sense (5'-GAG GCA TAA CTA TAG GAG ATT GGA AG-3') and antisense (5'-CTA TCC TTT CCA AGT CTG AAC TGG-3') primers were used plus a housekeeping gene, histidyl tRNA synthetase (sense: 5'-CTT CAG GGA GAG CGC GTG CG-3'; antisense: 5'-CCT TCA GGT CAT AGA TAA GC-3').…”

Section: To the Editormentioning

confidence: 99%

“…RT-PCR conditions were 94°C for 2 min, 30 cycles of denaturation at 94°C for 30 s, annealing at 58°C for 30 s, and extension at 72°C for 1 min (plus a final extension of 6 min at 72°C). The system was validated using cDNA extracted from cell lines (BJAB and AG876) that harbored ERV-3 5 . The PCR amplicons (ERV-3 404 bp and HtRNA 319 bp) were analyzed following gel electrophoresis by measuring their pixel intensities using Scion image software (Scion, Frederick, MD, USA).…”

Section: To the Editormentioning

confidence: 99%

See 2 more Smart Citations

Endogenous Retrovirus Erv-3 Is Not Implicated in Rheumatoid Arthritis But May Provide a Biomarker for Osteoarthritis: Table 1

Nelson¹,

Davari-Ejtehadi²,

Nevill³

et al. 2010

J Rheumatol

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The Rec protein of HERV‐K(HML‐2) upregulates androgen receptor activity by binding to the human small glutamine‐rich tetratricopeptide repeat protein (hSGT)

Hanke

Chudak

Kurth

et al. 2012

Intl Journal of Cancer

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The expression of endogenous retroviruses of the HERV‐K(HML‐2) family is strongly upregulated in germ cell tumors and several other cancers. Although the accessory Rec protein of HERV‐K(HML‐2) has been shown to induce carcinoma in situ in transgenic mice, to increase the activity of c‐myc and to interact with the androgen receptor (AR), whether or not Rec expression is indeed implicated causally in the initiation or progression of any human malignancies remains unclear. We used the yeast two‐hybrid system involving the Rec protein of a recently integrated HERV‐K(HML‐2) element in an effort to identify potential Rec‐related oncogenic mechanisms. This revealed the human small glutamine‐rich tetratricopeptide repeat (TPR)‐containing protein (hSGT) to be a cellular binding partner. The interaction of Rec with this known negative regulator of the AR was confirmed by coimmunoprecipitation, pull‐down assays and colocalization studies. The interaction involves the TPR motif of hSGT and takes place in the cytoplasm and in the nucleoli. Using an AR‐responsive promoter and gene we could demonstrate that Rec interference with hSGT resulted in an up to five‐fold increase in the activity of AR. Furthermore, in AR positive cells, Rec was shown to act as transactivator by enhancing AR‐mediated activation of the HERV‐K(HML‐2) LTR promoter. This is in line with previous observations of elevated HERV‐K(HML‐2) expression in steroid‐regulated tissues. On the basis of our findings we propose a “vicious cycle” model of Rec‐driven hyperactivation of the AR leading to increased cell proliferation, inhibition of apoptosis and eventually to tumor induction or promotion.

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A novel multiplex RT-PCR system detects human endogenous retrovirus-K in breast cancer

Cited by 23 publications

References 25 publications

Human Endogenous Retrovirus Type K (HERV-K) Particles Package and Transmit HERV-K–Related Sequences

Human Endogenous Retrovirus Type K (HERV-K) Particles Package and Transmit HERV-K–Related Sequences

Endogenous Retrovirus Erv-3 Is Not Implicated in Rheumatoid Arthritis But May Provide a Biomarker for Osteoarthritis: Table 1

The Rec protein of HERV‐K(HML‐2) upregulates androgen receptor activity by binding to the human small glutamine‐rich tetratricopeptide repeat protein (hSGT)

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