부산대학교 도서관

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.jmb.2006.07.085 Byline: Alicia Perez-Gonzalez, Ariel Rodriguez, Maite Huarte, IA[+ or -]igo J. Salanueva, Amelia Nieto Keywords: hCLE/CGI-99; transcription; RNAP II; influenza virus; CTD-phosphorylation Abbreviations: RNAP, RNA polymerase; RNP, ribonucleoprotein; CTD, C-terminal domain; HPRI, human placental ribonuclease inhibitor Abstract: The human protein hCLE was previously identified by its interaction with the PA subunit of influenza virus polymerase. It exhibits a sequence similarity of 38% with the yeast Spt16 component of the FACT complex, which is involved in transcriptional regulation. Therefore, we studied the possible relationship of hCLE with the transcription machinery. Here we show that hCLE and different phosphorylated forms of the RNA polymerase II (RNAP II) largest subunit, co-immunoprecipitate and colocalize by confocal microscopy analysis. Furthermore, hCLE was found in nuclear sites of active mRNA synthesis, as demonstrated by its colocalization with spots of in situ Br-UTP incorporation. Silencing of hCLE expression by RNA interference inhibited the synthesis of RNAP II transcripts around 50%. Accordingly, the expression profiling in hCLE-silenced cells studied by microarray analysis showed that, among the genes that exhibited a differential expression under hCLE silencing, more than 90% were down-regulated. Collectively these results indicate that hCLE works as a positive modulator of the RNA polymerase II activity. Author Affiliation: Centro Nacional de Biotecnologia, Cantoblanco, 28049 Madrid, Spain Article History: Received 12 May 2006; Revised 28 July 2006; Accepted 31 July 2006 Article Note: (miscellaneous) Edited by J. Karn