Abstract
Translocated in liposarcoma (TLS)/fused in sarcoma (FUS) is a multitasking DNA/RNA binding protein implicated in cancer and neurodegenerative diseases. Upon DNA damage, TLS is recruited to the upstream region of the cyclin D1 gene (CCND1) through binding to the promotor associated non-coding RNA (pncRNA) that is transcribed from and tethered at the upstream region. Binding to pncRNA is hypothesized to cause the conformational change of TLS that enables its inhibitive interaction with histone acetyltransferases and resultant repression of CCND1 expression, although no experimental proof has been obtained. Here, the closed-to-open conformational change of TLS on binding pncRNA was implied by fluorescence resonance energy transfer. A small fragment (31 nucleotides) of the full-length pncRNA (602 nucleotides) was shown to be sufficient for the conformational change of TLS. Dissection of pncRNA identified the G-rich RNA sequence that is critical for the conformational change. The length of RNA was also revealed to be critical for the conformational change. Furthermore, it was demonstrated that the conformational change of TLS is caused by another target DNA and RNA, telomeric DNA and telomeric repeat-containing RNA. The conformational change of TLS on binding target RNA/DNA is suggested to be essential for biological functions.
Highlights
Translocated in liposarcoma (TLS)/fused in sarcoma (FUS) is a multitasking DNA/RNA binding protein implicated in cancer and neurodegenerative diseases
blue fluorescent protein (BFP)-TLS-green fluorescent protein (GFP)-6xHis protein was successfully expressed in E. coli cells and purified by Ni-affinity chromatography followed by size-exclusion chromatography (SEC)
The conformational change of TLS caused by binding of the full-length promotor associated non-coding RNA (pncRNA) (602 nucleotides) was examined by fluorescence resonance energy transfer (FRET)
Summary
Translocated in liposarcoma (TLS)/fused in sarcoma (FUS) is a multitasking DNA/RNA binding protein implicated in cancer and neurodegenerative diseases. TLS is reportedly involved in shortening of the telomere, which is located at the end of the human chromosome, through repression of telomerase-independent telomere-elongation[12] In this biological process, TLS was found to be included in the ternary complex with telomeric DNA and telomeric repeat-containing RNA (TERRA), whose sequences are d(TTAGGG)n and r(UUAGGG)n, respectively. It seems important to investigate the relative roles of the different RNA binding domains and other regions of TLS in the context of the full-length protein to understand the molecular mechanism by which TLS regulates various different biological processes
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