title: Inhibition of human cytomegalovirus replication by overexpression of CREB1.
abstract: Expression of the human cytomegalovirus (HCMV) major immediate-early (MIE) genes is regulated by a strong enhancer-containing promoter with multiple binding sites for various transcription factors, including cyclic AMP response element binding protein 1 (CREB1). Here we show that overexpression of CREB1 potently blocked MIE transcription and HCMV replication. Surprisingly, CREB1 still exhibited strong inhibition of the MIE promoter when all five CREB binding sites within the enhancer were mutated, suggesting that CREB1 regulated the MIE gene expression indirectly. Promoter deletion analysis and site-directed mutagenesis identified the region between -130 and -50 upstream of the transcription start site of the MIE gene as the "CREB1 responsive region". Mutations of SP1/3 and NF-κB binding sites within this region interrupted the inhibitory effect induced by CREB1 overexpression. Our findings suggest that overexpression of CREB1 can cause repression of HCMV replication and may contribute to the development of new anti-HCMV strategies.
authors: Expression Engineering, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A(∗)STAR), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore.
authors: Yi Ling
authors: YL
authors: Chia
authors: Expression Engineering, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A(∗)STAR), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore.
authors: Chew Har
authors: CH
authors: Ng
authors: Department of Microbiology, University of Iowa, 51 Newton Rd., 3-772 Bowen Science Building, Iowa City, IA 52242, USA.
authors: Philip
authors: P
authors: Lashmit
authors: Expression Engineering, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A(∗)STAR), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore.
authors: Kai Ling
authors: KL
authors: Chu
authors: Expression Engineering, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A(∗)STAR), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore.
authors: Qiao Jing
authors: QJ
authors: Lew
authors: Expression Engineering, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A(∗)STAR), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore.
authors: Jia Pei
authors: JP
authors: Ho
authors: Microarray Groups, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A(∗)STAR), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore.
authors: Hsueh Lee
authors: HL
authors: Lim
authors: Microarray Groups, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A(∗)STAR), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore.
authors: Peter Morin
authors: PM
authors: Nissom
authors: Department of Microbiology, University of Iowa, 51 Newton Rd., 3-772 Bowen Science Building, Iowa City, IA 52242, USA.
authors: Mark F
authors: MF
authors: Stinski
authors: Expression Engineering, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A(∗)STAR), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore; Department of Microbiology, National University of Singapore, Block MD4, 5 Science Drive 2, Singapore 117597, Singapore. Electronic address: jimmy_chao@bti.a-star.edu.sg.
authors: Sheng-Hao
authors: SH
authors: Chao
mesh_terms: D002460:Cell Line; D017362:Cyclic AMP Response Element-Binding Protein / Q000096:biosynthesis*; D003587:Cytomegalovirus / Q000235:genetics / Q000502:physiology*; D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000235:genetics; D015870:Gene Expression; D017781:Genes, Immediate-Early; D054884:Host-Pathogen Interactions; D006801:Humans; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D014779:Virus Replication
chemical_list: C115163:CREB1 protein, human; D017362:Cyclic AMP Response Element-Binding Protein; D004279:DNA, Viral
keywords: CREB1; Human cytomegalovirus; Major immediate-early genes; NF-κB; SP1; SP3