Tags

Type your tag names separated by a space and hit enter

Multiple mechanisms are implicated in the regulation of NF-kappa B activity during human cytomegalovirus infection.
Proc Natl Acad Sci U S A. 1993 Feb 01; 90(3):1107-11.PN

Abstract

Infection-induced activation of the human cytomegalovirus major immediate early enhancer/promoter has been shown to be regulated primarily by transcription factor NF-kappa B cis elements. However, the mechanism(s) by which human cytomegalovirus induces NF-kappa B activity is unknown. A study was therefore undertaken to determine how this virus would affect normal NF-kappa B regulation. Viral infection of fibroblasts resulted in the specific stimulation of promoters containing major histocompatibility complex NF-kappa B cis elements fused upstream of the chloramphenicol acetyltransferase reporter gene. Electrophoretic mobility shift assays of nuclear extracts derived from mock- and virus-infected cells showed dramatic and sustained increases in DNA-binding proteins specific for these NF-kappa B sequences. Experiments using MAD-3 I kappa B, a specific inhibitor of NF-kappa B, and antibodies directed against rel family members demonstrated that the induced binding activities contained p50 and p65 proteins but not c-rel. Northern analysis indicated maximal levels of p50 mRNA by 4 h postinfection, whereas p65 and MAD-3 I kappa B mRNA accumulation peaked at 48-72 h postinfection, suggesting different regulatory mechanisms for p50 and p65/I kappa B genes. Electrophoretic mobility shift assays with deoxycholate-treated cytoplasmic extracts demonstrated a 3- to 4-fold decrease in the cytosolic stores of NF-kappa B binding activity by 4 h postinfection. Western blots probed with antibodies directed against MAD-3 I kappa B or pp40 (a protein isolated from chicken with sequence and biochemical properties similar to those of MAD-3 I kappa B) indicated that a cross-reactive peptide of 39 kDa was no longer detectable after 24 h postinfection. These results demonstrate that the activation and maintenance of nuclear NF-kappa B DNA binding and enhancer activities upon human cytomegalovirus infection occurs by multiple mechanisms.

Authors+Show Affiliations

Lineberger Comprehensive Cancer Center, School of Medicine, University of North Carolina, Chapel Hill 27599-7295.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

8381532

Citation

Kowalik, T F., et al. "Multiple Mechanisms Are Implicated in the Regulation of NF-kappa B Activity During Human Cytomegalovirus Infection." Proceedings of the National Academy of Sciences of the United States of America, vol. 90, no. 3, 1993, pp. 1107-11.
Kowalik TF, Wing B, Haskill JS, et al. Multiple mechanisms are implicated in the regulation of NF-kappa B activity during human cytomegalovirus infection. Proc Natl Acad Sci U S A. 1993;90(3):1107-11.
Kowalik, T. F., Wing, B., Haskill, J. S., Azizkhan, J. C., Baldwin, A. S., & Huang, E. S. (1993). Multiple mechanisms are implicated in the regulation of NF-kappa B activity during human cytomegalovirus infection. Proceedings of the National Academy of Sciences of the United States of America, 90(3), 1107-11.
Kowalik TF, et al. Multiple Mechanisms Are Implicated in the Regulation of NF-kappa B Activity During Human Cytomegalovirus Infection. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):1107-11. PubMed PMID: 8381532.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Multiple mechanisms are implicated in the regulation of NF-kappa B activity during human cytomegalovirus infection. AU - Kowalik,T F, AU - Wing,B, AU - Haskill,J S, AU - Azizkhan,J C, AU - Baldwin,A S,Jr AU - Huang,E S, PY - 1993/2/1/pubmed PY - 2001/3/28/medline PY - 1993/2/1/entrez SP - 1107 EP - 11 JF - Proceedings of the National Academy of Sciences of the United States of America JO - Proc Natl Acad Sci U S A VL - 90 IS - 3 N2 - Infection-induced activation of the human cytomegalovirus major immediate early enhancer/promoter has been shown to be regulated primarily by transcription factor NF-kappa B cis elements. However, the mechanism(s) by which human cytomegalovirus induces NF-kappa B activity is unknown. A study was therefore undertaken to determine how this virus would affect normal NF-kappa B regulation. Viral infection of fibroblasts resulted in the specific stimulation of promoters containing major histocompatibility complex NF-kappa B cis elements fused upstream of the chloramphenicol acetyltransferase reporter gene. Electrophoretic mobility shift assays of nuclear extracts derived from mock- and virus-infected cells showed dramatic and sustained increases in DNA-binding proteins specific for these NF-kappa B sequences. Experiments using MAD-3 I kappa B, a specific inhibitor of NF-kappa B, and antibodies directed against rel family members demonstrated that the induced binding activities contained p50 and p65 proteins but not c-rel. Northern analysis indicated maximal levels of p50 mRNA by 4 h postinfection, whereas p65 and MAD-3 I kappa B mRNA accumulation peaked at 48-72 h postinfection, suggesting different regulatory mechanisms for p50 and p65/I kappa B genes. Electrophoretic mobility shift assays with deoxycholate-treated cytoplasmic extracts demonstrated a 3- to 4-fold decrease in the cytosolic stores of NF-kappa B binding activity by 4 h postinfection. Western blots probed with antibodies directed against MAD-3 I kappa B or pp40 (a protein isolated from chicken with sequence and biochemical properties similar to those of MAD-3 I kappa B) indicated that a cross-reactive peptide of 39 kDa was no longer detectable after 24 h postinfection. These results demonstrate that the activation and maintenance of nuclear NF-kappa B DNA binding and enhancer activities upon human cytomegalovirus infection occurs by multiple mechanisms. SN - 0027-8424 UR - https://www.unboundmedicine.com/medline/citation/8381532/Multiple_mechanisms_are_implicated_in_the_regulation_of_NF_kappa_B_activity_during_human_cytomegalovirus_infection_ L2 - http://www.pnas.org/cgi/pmidlookup?view=long&pmid=8381532 DB - PRIME DP - Unbound Medicine ER -