The glass transition temperature (Tg) of polymer nanostructures was measured using a technique based on synchrotron x-ray diffraction from periodic grating structures. Poly(methyl methacrylate) (PMMA) nanostructures consisting of 1:1 lines:spaces with a 100 nm period and 100 nm height were characterized to have a Tg of 118°C, which is comparable to the Tg of PMMA in bulk systems. The Tg of the PMMA structures also was measured as a function of absorbed x-ray dose. Doses ranging from 0 to 2400 mJ/mm3 were delivered to the PMMA structures prior to the Tg measurements; the Tg of the structures was found to decrease from 118°C to 95°C, respectively. The dose dependence of the PMMA glass transition temperature can be attributed to changes in the polymer molecular weight under exposure to x rays.