+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Defective growth correlates with reduced accumulation of a viral DNA replication protein after low-multiplicity infection by a human cytomegalovirus ie1 mutant



Defective growth correlates with reduced accumulation of a viral DNA replication protein after low-multiplicity infection by a human cytomegalovirus ie1 mutant



Journal of Virology 72(1): 366-379



To investigate the importance of the IE1 p72 regulatory protein during human cytomegalovirus replication, a recombinant virus unable to synthesize IE1 p72 was constructed. The Towne strain mutant CR208 lacked exon 4 of the major immediate-early gene and was isolated and complemented in an IE1-expressing immortalized human fibroblast line (ihfie1.3). Replication of CR208 in primary human fibroblasts was completed after an input multiplicity of 10 PFU/cell but was severely impaired at 0.1 PFU/cell. CR208 formed plaques with lower efficiency on primary fibroblasts than on ihfie1.3 cells, and the relationship between the CR208 inoculum size and the resulting number of undersized plaques was nonlinear, indicating that multiple particles of CR208 were required to initiate lytic replication in a single primary fibroblast. After infection of primary fibroblasts with CR208 at 5 PFU/cell, a normal pattern of viral antigens was detected, although IE1 p72 was absent. During lower-multiplicity infections, IE2 protein was consistently detected at similar levels in a similar proportion of CR208-infected cells relative to the case for a Towne infection, but many fewer CR208-infected cells contained the ppUL44 polymerase accessory protein when evaluated at 24 or 48 h after infection. Furthermore, fibroblasts infected with CR208 at a low multiplicity failed to form viral DNA replication compartments, despite having expressed IE2 p86. These low-multiplicity growth and expression defects were corrected in two rescued derivatives of CR208 able to synthesize IE1 p72. One rescued virus (CR249) carried a deletion removing the large intron between exons 1 and 2 of the ie1-ie2 locus, revealing that this intron was dispensable for growth in cell culture.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 008423373

Download citation: RISBibTeXText

PMID: 9420235


Related references

Processing bodies accumulate in human cytomegalovirus-infected cells and do not affect viral replication at high multiplicity of infection. Virology 458-459: 151-161, 2014

Human cytomegalovirus UL84 insertion mutant defective for viral DNA synthesis and growth. Journal of Virology 78(19): 10360-9, 2004

The human cytomegalovirus major immediate-early enhancer determines the efficiency of immediate-early gene transcription and viral replication in permissive cells at low multiplicity of infection. Journal of Virology 77(6): 3602-3614, 2003

Disruption of PML-associated nuclear bodies by IE1 correlates with efficient early stages of viral gene expression and DNA replication in human cytomegalovirus infection. Virology 274(1): 39-55, 2000

Absence of IE1 p72 protein function during low-multiplicity infection by human cytomegalovirus results in a broad block to viral delayed-early gene expression. Journal of Virology 76(9): 4441-4455, 2002

Enhanced viral pathogenesis associated with a virulent mutant virus or a virulent satellite RNA correlates with reduced virion accumulation and abundance of free coat protein. Virology 312(1): 8-13, 2003

Replication of human cytomegalovirus at supra-optimal temperatures is dependent on the virus strain, multiplicity of infection and phase of virus replication. Journal of General Virology 51(Pt 1): 83-97, 1980

Restriction of the replication of human cytomegalovirus at supraoptimal temperature is dependent on the virus strain multiplicity of infection and phase of virus replication. Nelson, J D And C Grassi (Ed ) Current Chemotherapy And Infectious Disease, Vol 1 And 2; Proceedings Of The 11th International Congress And The 19th Interscience Conference on Antimicrobial Agents And Chemotherapy, Boston, Mass , Usa, Oct 1-5, 1979 Xxi+774p (Vol 1); Ix+1023p (Vol 2 ) The American Society For Microbiology: Washington, D C , Usa Illus P1314-1316, 1980

Transfer of the UAP56 Interaction Motif of Human Cytomegalovirus pUL69 to Its Murine Cytomegalovirus Homolog Converts the Protein into a Functional mRNA Export Factor That Can Substitute for pUL69 during Viral Infection Genome Replication and Regulation. 2012

Growth characteristics of cytomegalovirus in human fibroblasts with demonstration of protein synthesis early in viral replication. Journal of Virology 11(6): 991-997, 1973

A deletion mutant in the human cytomegalovirus gene encoding IE1(491aa) is replication defective due to a failure in autoregulation. Proceedings of the National Academy of Sciences of the United States of America 93(21): 11321-6, 1996

A replication-defective human cytomegalovirus vaccine for prevention of congenital infection. Science Translational Medicine 8(362): 362ra145, 2016

Small ubiquitin-related modifier (SUMO) pathway-mediated enhancement of human cytomegalovirus replication correlates with a recruitment of SUMO-1/3 proteins to viral replication compartments. Journal of General Virology 94(Pt 6): 1373-1384, 2013

Daxx-mediated accumulation of human cytomegalovirus tegument protein pp71 at ND10 facilitates initiation of viral infection at these nuclear domains. Journal of Virology 76(15): 7705-7712, 2002