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Molecular Biology Faculty

S. Jane Flint

Professor of Molecular Biology

Jane Flint

 

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Phone (609) 258-6113
locationLewis Thomas Lab, 234
Phone Lab (609) 258-5414
Faculty Assistant
Ellen Brindle-Clark
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Phone (609) 258-5419

Research Focus

Modulation of host cell processes by adenoviral gene products

Human adenoviruses, which are widespread in the population, are non-enveloped viruses with double-stranded DNA genomes. The study of these viruses has yielded fundamental insights into the mechanisms by which genes are expressed in human cells, as well as the intricate circuits that control cell proliferation but become deranged during malignant transformation. There is considerable interest in developing derivatives of adenoviruses for therapeutic applications, for example, as gene transfer vectors and oncolytic viruses for cancer therapy. However, relatively little attention has been paid to the interactions among viral and cellular gene products required for optimal virus reproduction in normal human cells (as opposed to highly abnormally transformed cells derived from human tumors). Furthermore, the molecular functions of the products of genes routinely deleted from “therapeutic” adenoviruses are not fully understood. Our research seeks to address such issues, focusing on the contributions of the viral E1B 55 kDa protein to the infectious cycle in normal human cells: the gene for this protein is deleted from all adenovirus gene transfer vectors, and such deletion (in the absence of other alterations) confers tumor cell-selective reproduction and host cell lysis.

One function of this E1B protein implicated in the cell-selective reproduction is induction of selective export of viral mRNAs during the late phase of infection. This process requires assembly in infected cells of a virus-specific E3 ubiquitin ligase that contains the viral E1B 55 kDa and E4 Orf6 proteins, as well as several cellular proteins. We have established that this virus-specific enzyme targets one or more components of the major mRNA export pathway in mammalian cells, the Nxf1 pathway, and that efficient export of viral late mRNAs is more dependent on the E1B 55 kDa protein in normal than in tumor cells. As the relevant substrates of the virus specific E3 ubiquitin ligase have not been identified, we are applying proteomic methods to compare the populations of ubiquitinylated proteins in cells infected by mutant viruses defective for production of the E1B 55 kDa protein and their wild-type parent.

One of the first activities ascribed to the E1B 55 kDa pote9in was repression of transcription in simplified experimental systems, such as transient expression assays. However, the contribution of this function to viral reproduction was not known, until the results of our genome-wide analysis of cellular gene expression in infected normal human cells established that the E1B protein inhibits expression of a substantial number of genes. This set of genes proved to highly enriched for those associated with anti-viral defenses, particularly that mediated by the cytokine type I interferon. In fact, the E1B 55 kDa protein is required to block inhibition of viral reproduction by interferon, and when the E1B protein is not made, viral genome replication is inhibited severely in infected cells exposed to the interferon. The results of molecular and genetic analysis have established that this viral protein represses transcription of interferon-inducible genes and that his function correlates with preventing inhibition of viral DNA synthesis by interferon. This cytokine does not induce inhibition of synthesis of viral replication proteins, but rather leads to a block in formation of the specialized intranuclear structures in which viral DNA synthesis takes place, so-called viral replication centers. The mechanism by which the E1B 55 kDa protein represses transcription of specific human genes is currently under investigation, as is that by which the products of one or more of the some 130 human genes that are both interferon-inducible and E1B protein repressed disrupt formation of viral replication centers.


Selected Publications

Dehart CJ, Chahal JS, Flint SJ, Perlman DH. (2013) Extensive post-translational modification of active and inactivated forms of endogenous p53. Mol Cell Proteomics. 13: 1-17.  PubMed

Chahal JS, Flint SJ. (2013) The p53 protein does not facilitate adenovirus type 5 replication in normal human cells. J Virol. 87: 6044-46. PubMed

Chahal JS, Gallagher C, Dehart CJ, Flint SJ. (2013) The repression domain of the E1B 55 kDa protein participates in countering interferon-induced inhibition of adenovirus replication. J Virol. 87: 4432-44. PubMed

Kato SE, Chahal JS, Flint SJ. (2012) Reduced infectivity of adenovirus type 5 particles and degradation of entering viral genomes associated with incomplete processing of the pre-terminal protein. J Virol. 86: 13554-65. PubMed

Chahal JS, Qi J, Flint SJ. (2012) The human adenovirus type 5 E1B 55 kDa protein obstructs inhibition of viral replication by type I interferon in normal human cells. PLoS Pathog. 8: e1002853. PubMed

Kato SE, Huang W, Flint SJ. (2011) Role of the RNA recognition motif of the E1B 55 kDa protein in the adenovirus type 5 infectious cycle.  Virology 417(1):9-17.  PubMed

Miller DL, Rickards B, Mashiba M, Huang W, Flint SJ. (2009) The adenoviral E1B 55-kilodalton protein controls expression of immune response genes but not p53-dependent transcription. J Virol. 83: 3591-603. PubMed

Pérez-Berná AJ, Marabini R, Scheres SH, Menéndez-Conerjero R, Dmitriev IP, Curiel DT, Mangel WF, Flint SJ, San Martín C. (2009) Structure and uncoating of immature adenovirus.  J. Mol. Biol. 392(2):547-57.  PubMed

LeRoy G, Rickards B, Flint SJ. (2008) The double bromodomain proteins Brd2 and Brd3 couple histone acetylation to transcription. Mol Cell. 30: 51-60. PubMed

Miller DL, Myers CL, Rickards B, Coller HA, Flint SJ. (2007) Adenovirus type 5 exerts genome-wide control over cellular programs governing proliferation, quiescence, and survival. Genome Biol. 8: R58. PubMed

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