Ubiquitin-mediated proteolysis is an integral part of diverse cellular functions, and of the three enzymes involved in linking ubiquitin to protein targets, the E3 ubiquitin ligases are of particular interest as they confer substrate specificity during this process. The E3 ubiquitin ligases can be categorized based on mechanism of action and on the presence of specific domains such as RING, HECT, F-box, and U-box. In plants, the U-box family has undergone a large gene expansion that may be attributable to biological processes unique to the plant life cycle. For example, there are 64 predicted plant U-box (PUB) proteins in Arabidopsis, and the biological roles of many of these have yet to be determined. Research on PUB genes from several different plants has started to elucidate a range of functions for this family, from self-incompatibility and hormone responses to defence and abiotic stress responses. Expression profiling has also been used as a starting point to elucidate PUB function, and has uncovered a strong connection of PUB genes to various stress responses. Finally, some PUB proteins have been linked to receptor kinases as upstream activators, and downstream target substrates are also starting to emerge. The mechanisms of action range from the observation of mono-ubiquitination during non-proteolytic signalling to directed regulation of proteasomal components during stress responses, and cell death appears to be a theme underlying many PUB functions.