RNA polymerase III control elements are required for trans-activation by the murine retroviral long terminal repeat sequences

Yong Woo Hwang, Na Kyung Yoo, Heung Mo Yang, Sang-Yun Choi

Research output: Contribution to journalArticle

Abstract

RNA leukemia viruses induce T-cell lymphoblastic lymphomas or myeloid leukemias. Infection of cells with Moloney murine leukemia virus (M-MuLV) up-regulates the expression of a number of cellular genes, including those involved in T-lymphocyte activation. Previously, we demonstrated that this up-regulation occurs via the trans-activation activity of the M-MuLV long terminal repeat (LTR) sequences which produce an LTR-encoded transcript. Sequence analysis of the LTR revealed a potential transcription unit for RNA polymerase III (Pol III) within the U3 region that is actively occupied by Pol II factors. Here, we provide the direct evidence of involvement of Pol III in the trans-activation process and demonstrate the precise localization of the intragenic control elements for accurate and active Pol III transcription. Deletions of a copy of the directed repeats and further immediate upstream sequences significantly abrogated the generation of LTR-encoded transcript and abolished the trans-activational activity, whereas the deletion of a copy of directed repeats alone proportionally reduced the transcript size, but still retained moderately high trans-activational activity. In electrophoretic mobility shift assay, the fraction containing a multiple transcription factor TFIIIC complex strongly bound to the LTR-U3 probe containing the essential control elements. The specificity of the DNA-TFIIIC interaction was confirmed by conducting competition assays with DNA fragments containing a genuine Pol III-transcribed gene, VA1, and by vaccinia virus infection which stimulates the expression of Pol III factors. However, a deletion mutant lacking an essential control element bound to the TFIIIC complex poorly, consequently resulting in weak Pol III transcription as assessed by an IRES-GFP reporter system. This correlation strongly supports the possibility that the generation of LTR-encoded transcript is directed by Pol III. Therefore, this finding suggests the involvement of Pol III transcription in the retrovirus-induced activation of cellular genes, potentially contributing to leukemogenesis.

Original languageEnglish
Pages (from-to)110-115
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume456
Issue number1
DOIs
Publication statusPublished - 2015 Jan 2

Fingerprint

RNA Polymerase III
Terminal Repeat Sequences
Chemical activation
Transcription
Viruses
Moloney murine leukemia virus
T-cells
Genes
Precursor Cell Lymphoblastic Leukemia-Lymphoma
Assays
Up-Regulation
T-Cell Leukemia
Electrophoretic mobility
Myeloid Leukemia
T-Cell Lymphoma
Vaccinia virus
RNA Viruses
DNA
Electrophoretic Mobility Shift Assay
Virus Diseases

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

RNA polymerase III control elements are required for trans-activation by the murine retroviral long terminal repeat sequences. / Hwang, Yong Woo; Yoo, Na Kyung; Yang, Heung Mo; Choi, Sang-Yun.

In: Biochemical and Biophysical Research Communications, Vol. 456, No. 1, 02.01.2015, p. 110-115.

Research output: Contribution to journalArticle

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