Apple ships some configurations with only three slots populated, so empty slot ARE permitted. [I believe you can run ONE 16GB stick, but no one seems to have reported testing that configuration.]
Experts, including OWC [lllaass has provided the link to their study above] have determined that sizes can be freely mixed EXCEPT: 16GB modules are Registered (contain a buffer register) and cannot be intermixed with Unregistered smaller modules. [so if you ever anticipate going to multiple 16GB modules, consider starting now.]
There are slight speed improvements when size-and-speed-matched modules are paired in adjacent slots, but real-world speed differences appear to be very small indeed. (this is probably due the the multiple levels of caches used in the Xeon Processors.)
When buying modules for Macs, always buy from a Mac-Centric vendor who assures you:
"It works in your Mac,
or your money back."
Best installation practice is to test the new modules by themselves to assure they are working, THEN add old modules to decide whether old and new can "play nice together".
Remember that Mac Pro with Xeon Processor has error-correcting memory. Problems do not fester undetected -- real issues will be immediately obvious. (such as the slots will be declared "empty" or the Mac will just sit there and blink the power light.)