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Su, Justin - Assignment #3 - IMRAD
Su, Justin - Assignment #3 - IMRAD
Justin Su
Doug Bradley
WRIT 159A – Winter 2019
February 5, 2019
I have chosen a published article from the Journal of Virology that was published on December
13, 2017. “Upon Infection, Cellular WD Repeat-Containing Protein 5 (WDR5) Localizes to
Cytoplasmic Inclusion Bodies and Enhances Measles Virus Replication” was authored by
Dzwokai Ma, Cyril X. George, Jason L. Nomburg, Christian K. Pfaller, Roberto Cattaneo, and
Charles E. Samuel. The structure of this article follows the Introduction, Results, Discussion, and
Methods (IRDAM) format. This article analysis serves to identify and summarize each IRDAM
section.
Introduction
The introduction of the article begins describing the known measles virus (MeV) proteins and their
functions. Containing single-stranded negative-sense RNA genome of ∼15.9 kilobases, the MeV
capsid is composed of the nucleocapsid (N) protein, which would contain the L (large) and P
(phospho) MeV proteins. According to an electron microscopic (EM) study, these proteins are
known to form inclusion bodies within HeLa cells. In addition, heat shock protein 70 (HSP 70)
was also found to interact with the inclusion bodies.
What was the aim of the study? What hypothesis did the researchers test? Are the conclusions
reached (assuming they are valid) important to you and others? (Explain.)
The aim of the study is to see whether WDR5, a regulatory subunit of histone 3 lysine 4
methyltransferases (H3K4MTs), influences MeV replication. The author’s hypothesis was based
on preliminary data, in which WDR5-EGFP was found localized to the inclusion bodies. The
conclusion of this hypothesis will shed light on a mechanism of cytoplasmic functions for the
subunits of H3K4MTs. Initiatives on this study may provide implications on the pathology of
measles virus, which is currently plaguing the State of Washington.
Results
The results of this article extensively outlined the outcomes of each experiment performed. The
first experiment continued the exposition that led to the hypothesis. WDR5-EGFP was found
localized to cytoplasmic puncta of MeV infected HeLa cells, in which EGFP was not. Therefore,
it was clear through fluorescent live-cell imaging to not only avoid artifacts from fixation, but also
reveals the total endogenous WDR5-EGFP on these puncta. Another experiment investigated
whether other subunits of H3K4MTs were found localized to these puncta, which was not evident
through fluorescent live-cell imaging. Furthermore, the authors performed experiments to test if
MeV proteins were associated with WDR5-EGFP. Immunoprecipitation results were shown,
indicating that MeV L, P, N, and C complexes with WDR5-EGFP. In addition, a siRNA
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knockdown was performed on WDR5, resulting in reduced MeV protein expression. This implies
that without WDR5, MeV replication was reduced. Lastly, MeV infection did not lead to a decrease
or increase in H3K4me3 as shown in Figure 9.
Discussion
The discussion emphasized the importance of WDR5 as a host factor in measles virus, where host-
pathogen relationships are key in studying the measles viral pathology. The authors summarized
their results, and discussed further implications of their study. They considered alternative
hypotheses and went over why the rationales of other hypotheses are not likely. They have
suggested that their future studies can elucidate the formation of the viral RNA replication
complex, nucleocapsid trafficking, and shielding from host innate immune response sensors.
Methods
The methods used in this article were detailed and written in such a way that the experiments may
be reproduced in their entirety. The cell line, viruses, RNAi, and the procedures of the Western
Blotting, immunoprecipitation, and histone extraction were all listed in detail. The equipment used,
such as the microscope models, were listed as well.
Was the statistical analysis (if used) appropriate for the study?
The student’s t-test on Microsoft Excel was used to acquire the significance of the Western Blots,
where the null hypothesis was rejected when the p value was less than 5%.
Acknowledgements
The acknowledgements highlighted the distribution of effort in publishing this article, listing who
developed the experiments and who conducted the experiments.