Characterization of 3′ termini of human cytomegalovirus UL138-UL145 transcripts in a clinical strain

Ma, Yanping; He, Rong; Wang, Ning; Ruan, Qiang; Ji, Yaohua; Li, Mali; Sun, Zhengrong; Ren, Gaowei

doi:10.1111/j.1348-0421.2010.00294.x

Cited by 12 publications

(8 citation statements)

References 21 publications

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“…The relative conservation of UL145 sequences being retained under selection pressure suggests that it might be essential for virus survival and replication in vivo. Qi et al [2011] have identified the transcripts of the UL138-UL145 region as two large families of polycistronic transcripts, with the UL145 gene representing one component of the second family. He et al [2011] have confirmed that UL144 and UL145 are expressed as a polycistron.…”

Section: Discussionmentioning

confidence: 99%

Transcriptional features and transcript structure of UL145 in different strains of human cytomegalovirus

Wang

Sun

et al. 2011

Journal of Medical Virology

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The human cytomegalovirus UL145 gene is located between the highly variable genes UL144 and UL146 in the UL/b' region. However, unlike its neighboring genes, UL145 is relatively conserved among strains isolated from patients. The transcriptional features and transcript structure of UL145 has not yet been examined. In this study, the transcriptional features and structure of UL145 were characterized using RNA preparations from three HCMV strains isolated from patients, designated H, C, and X, respectively. Two transcripts were identified by cDNA library screening. Two main clusters of transcripts, one located between 500 and 700 nt, and the other between 1,400 and 1,700 nt, were confirmed by Northern blot analysis. Abundant transcripts were detected at 96 h post-infection by Northern blot, suggesting that UL145 was a late gene. The terminal sequences of transcripts obtained by 3' rapid amplification of cDNA ends demonstrated the presence of polyadenylation. Transcripts identified in the RNA of H, C, and X strains 96 h post-infection had the same 3' end but different 5' ends. In addition to polycistronic transcripts with upstream genes, UL145 could be transcribed as a single transcript during late strain infections.

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Section: Discussionmentioning

confidence: 99%

Transcriptional features and transcript structure of UL145 in different strains of human cytomegalovirus

Wang

Sun

et al. 2011

Journal of Medical Virology

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“…A pair of specific primers for the UL21.5 gene region (R-s3 and 5'R3) (Table I and Fig. 1) were designed and used to screen UL21.5 genespecific clones from the cDNA library by graded polymerase chain reaction (PCR) as described in previous studies (21,22). The PCR reaction conditions were as follows: 95˚C for 5 min, 30 cycles of 95˚C for 45 sec, 56˚C for 45 sec and 72˚C for 30 sec, followed by a final elongation at 72˚C for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Validation of three splice donor and three splice acceptor sites for regulating four novel low-abundance spliced transcripts of human cytomegalovirus UL21.5 gene locus

Gao

Ruan

et al. 2014

International Journal of Molecular Medicine

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In a previous study, one spliced transcript of human cytomegalovirus (HCMV), named UL21.5 was identified. UL21.5 has been found to be one of the viral transcripts packaged within HCMV particles. The UL21.5 mRNA is translated into a secreted glycoprotein, which is a viral chemokine decoy receptor specifically interacting with regulated upon activation normal T cell expressed and secreted (RANTES). In the present study, four novel low-abundance 3'-coterminal spliced transcripts were identified to be transcribed from the UL21.5 gene region of a low-passage HCMV strain during the late infection phase by cDNA library screening, northern blot hybridization, reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE)-PCR. Three splicing donor and three splicing acceptor sites found in the UL21.5 gene region were validated to be functional in an in vitro expression system. In addition, the determinant regulatory region that is necessary for the splice donor site at nucleotide (nt) 25533 was located in a 9-bp sequence around the site; the regulatory regions for the splice acceptor sites at nt 26597 and nt 26633 were located in a 20-bp sequence upstream of the site at nt 26597 and in a 10-bp sequence from nt 26641 to nt 26650 downstream of the site at nt 26633, respectively.

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“…To select specific cDNA clones from the cDNA library by PCR, a graded PCR was set up as previously described [19,20]. Five thousand cDNA clones were screened by graded PCR using several pairs of primers (Table 1, Figure 1).…”

Section: Methodsmentioning

confidence: 99%

Analysis and mapping of a 3′ coterminal transcription unit derived from human cytomegalovirus open reading frames UL30–UL32

Gao

Wang

et al. 2013

Virol J

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BackgroundIt has been predicted that the UL31 gene originates from the positive strand of the human cytomegalovirus (HCMV) genome, whereas the UL30 and UL32 genes originate from the complementary strand. Except for the UL32 gene, the transcription of this gene region has not been investigated extensively.MethodsNorthern blotting, cDNA library screening, RACE-PCR,and RT-PCR were used.ResultsAt least eight transcripts of the antisense orientation of UL31 were transcribed from the UL30–UL32 region during the late phase of HCMV infection. The 3′ coterminus of these transcripts was located within the predicted UL30 gene. The longest 6.0-kb transcript was initiated upstream of the predicted UL32 gene. Other transcripts were derived from the predicted UL30 and UL31 gene region. Except for the previously predicted UL32 open reading frame (ORF), three novel ORFs, named UL31anti-1, UL31anti-2 and UL31anti-3, were located in the transcripts from the UL31anti-UL32 transcription unit. No transcription was found in UL31.ConclusionA family of novel 3′ coterminal transcripts was transcribed from the UL30–UL32 gene region.

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Characterization of 3′ termini of human cytomegalovirus UL138-UL145 transcripts in a clinical strain

Cited by 12 publications

References 21 publications

Transcriptional features and transcript structure of UL145 in different strains of human cytomegalovirus

Transcriptional features and transcript structure of UL145 in different strains of human cytomegalovirus

Validation of three splice donor and three splice acceptor sites for regulating four novel low-abundance spliced transcripts of human cytomegalovirus UL21.5 gene locus

Analysis and mapping of a 3′ coterminal transcription unit derived from human cytomegalovirus open reading frames UL30–UL32

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