Abstract
Specific chromatin immunoprecipitation of salt-fractionated infected cell extracts has demonstrated that the CCCTC-binding factor (CTCF), a highly conserved, 11-zinc-finger DNA-binding protein with known roles in cellular and viral genome organization and gene expression, specifically binds the genome of Minute Virus of Mice (MVM). Mutations that diminish binding of CTCF to MVM affect processing of the P4-generated pre-mRNAs. These RNAs are spliced less efficiently to generate the R1 mRNA, and definition of the NS2-specific exon upstream of the small intron is reduced, leading to relatively less R2 and the generation of a novel exon-skipped product. These results suggest a model in which CTCF is required for proper engagement of the spliceosome at the MVM small intron and for the first steps of processing of the P4-generated pre-mRNA.
Highlights
Parvoviruses are small (20 nm) non-enveloped icosahedral viruses that infect and cause disease in many vertebrate hosts
We previously showed that Minute Virus of Mice (MVM) replicates in close association with sites on the cellular genome, taking advantage of the fact that these sites are replete with factors involved in gene expression and DNA damage signaling [13]
In surveying the MVM genome for the binding sites of known cellular factors, we noticed consensus CCCTC-binding factor (CTCF) binding sites in the NS and the VP regions of MVM that were conserved in a number of other parvoviruses
Summary
Parvoviruses are small (20 nm) non-enveloped icosahedral viruses that infect and cause disease in many vertebrate hosts. They are unique among all known animal viruses in that they contain ~5 kb single-stranded linear DNA genomes, with inverted terminal repeats at both ends, which form hairpin structures and serve as origins of replication [1]. Transcripts R3 (3.0 kb), generated from P38 promoter, encode the two viral capsid proteins, VP1 (83 kDa) and VP2 (64 kDa), utilizing the open reading frame (ORF) in the right half of the genome [4,5]. A small, overlapping intron, common to both P4- and P38-generated transcripts, utilizes two donors, D1 (nt 2280) and D2 (nt 2317) and two acceptors, A1 (nt 2377) and
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