Biology Faculty Articles
Title
Measurement of effector protein injection by type III and type IV secretion systems using a 13-residue phosphorylatable glycogen synthase kinase tag
Document Type
Article
Publication Date
1-1-2006
Publication Title
Infection and Immunity
ISSN
0019-9567
Volume
74
Issue/No.
10
First Page
5645
Last Page
5657
Abstract
Numerous bacterial pathogens use type III secretion systems (T3SSs) or T4SSs to inject or translocate virulence proteins into eukaryotic cells. Several different reporter systems have been developed to measure the translocation of these proteins. In this study, a peptide tag-based reporter system was developed and used to monitor the injection of T3S and T4S substrates. The glycogen synthase kinase (GSK) tag is a 13-residue phosphorylatable peptide tag derived from the human GSK-3beta kinase. Translocation of a GSK-tagged protein into a eukaryotic cell results in host cell protein kinase-dependent phosphorylation of the tag, which can be detected with phosphospecific GSK-3beta antibodies. A series of expression plasmids encoding Yop-GSK fusion proteins were constructed to evaluate the ability of the GSK tag to measure the injection of Yops by the Yersinia pestis T3SS. GSK-tagged YopE, YopH, LcrQ, YopK, YopN, and YopJ were efficiently phosphorylated when translocated into HeLa cells. Similarly, the injection of GSK-CagA by the Helicobacter pylori T4SS into different cell types was measured via phosphorylation of the GSK tag. The GSK tag provides a simple method to monitor the translocation of T3S and T4S substrates.
NSUWorks Citation
Garcia, Julie Torruellas; F. Ferracci; M. W. Jackson; S. S. Pattis; L.R.W. Plano; W. Fischer; and G. V. Plano. 2006. "Measurement of effector protein injection by type III and type IV secretion systems using a 13-residue phosphorylatable glycogen synthase kinase tag." Infection and Immunity 74, (10): 5645-5657. https://nsuworks.nova.edu/cnso_bio_facarticles/100
