Global human-kinase screening identifies therapeutic host targets against influenza

Colm Atkins, Carrie W. Evans, Brian Nordin, Matthew P. Patricelli, Robert Reynolds, Krister Wennerberg, James W. Noah

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

During viral infection of human cells, host kinases mediate signaling activities that are used by all viruses for replication; therefore, targeting of host kinases is of broad therapeutic interest. Here, host kinases were globally screened during human influenza virus (H1N1) infection to determine the time-dependent effects of virus infection and replication on kinase function. Desthiobiotin-labeled analogs of adenosine triphosphate and adenosine diphosphate were used to probe and covalently label host kinases in infected cell lysates, and probe affinity was determined. Using infected human A549 cells, we screened for time-dependent signal changes and identified host kinases whose probe affinities differed significantly when compared to uninfected cells. Our screen identified 10 novel host kinases that have not been previously shown to be involved with influenza virus replication, and we validated the functional importance of these novel kinases during infection using targeted small interfering RNAs (siRNAs). The effects of kinase-targeted siRNA knockdowns on replicating virus levels were measured by quantitative reverse-transcription PCR and cytoprotection assays. We identified several novel host kinases that, when knocked down, enhanced or reduced the viral load in cell culture. This preliminary work represents the first screen of the changing host kinome in influenza virus-infected human cells.

Original languageEnglish (US)
Pages (from-to)936-946
Number of pages11
JournalJournal of Biomolecular Screening
Volume19
Issue number6
DOIs
StatePublished - 2014
Externally publishedYes

Fingerprint

Human Influenza
Screening
Phosphotransferases
Viruses
Virus Diseases
Virus Replication
Orthomyxoviridae
Therapeutics
Small Interfering RNA
Cells
Cytoprotection
Transcription
Viral Load
Cell culture
Adenosine Diphosphate
Reverse Transcription
Labels
Assays
Cell Culture Techniques
Adenosine Triphosphate

Keywords

  • influenza
  • kinase
  • profiling
  • siRNA

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Pharmacology
  • Biochemistry
  • Molecular Medicine
  • Biotechnology

Cite this

Atkins, C., Evans, C. W., Nordin, B., Patricelli, M. P., Reynolds, R., Wennerberg, K., & Noah, J. W. (2014). Global human-kinase screening identifies therapeutic host targets against influenza. Journal of Biomolecular Screening, 19(6), 936-946. https://doi.org/10.1177/1087057113518068

Global human-kinase screening identifies therapeutic host targets against influenza. / Atkins, Colm; Evans, Carrie W.; Nordin, Brian; Patricelli, Matthew P.; Reynolds, Robert; Wennerberg, Krister; Noah, James W.

In: Journal of Biomolecular Screening, Vol. 19, No. 6, 2014, p. 936-946.

Research output: Contribution to journalArticle

Atkins, C, Evans, CW, Nordin, B, Patricelli, MP, Reynolds, R, Wennerberg, K & Noah, JW 2014, 'Global human-kinase screening identifies therapeutic host targets against influenza', Journal of Biomolecular Screening, vol. 19, no. 6, pp. 936-946. https://doi.org/10.1177/1087057113518068
Atkins, Colm ; Evans, Carrie W. ; Nordin, Brian ; Patricelli, Matthew P. ; Reynolds, Robert ; Wennerberg, Krister ; Noah, James W. / Global human-kinase screening identifies therapeutic host targets against influenza. In: Journal of Biomolecular Screening. 2014 ; Vol. 19, No. 6. pp. 936-946.
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