Mounting an effective innate immune response against pathogens requires the rapid and global reprogramming of host cellular processes. Here we employed complementary proteomic methods to identify differentially regulated proteins early during a plant's defense response. Besides defense-related proteins, constituents of the largest category of upregulated proteins were cytoplasmic- and ER-residing molecular chaperones. Investigating the significance of upregulated ER chaperones, we find that silencing of ER-resident protein disulfide isomerases NbERp57 and NbP5 and the calreticulins NbCRT2 and NbCRT3 led to partial loss of N immune receptor-mediated defense against Tobacco mosaic virus (TMV). Furthermore, NbCRT2 and NbCRT3 were required for the expression of a previously uncharacterized induced receptor-like kinase (IRK). IRK is a plasma membrane-localized protein required for N-mediated hypersensitive response, programmed cell death, and resistance to TMV. These data support a model in which ER-resident chaperones are required for the accumulation of membrane-bound or secreted proteins during plant innate immunity.