The victory sees Bernard Molekwa's charges end their five-game winless run in the South African topflight.
Rise and Shine broke the deadlock after 11 minutes courtesy of a thunderous free-kick from Ndivhuwo Ravhuhali that left Siyabonga Mbatha with no chance.
It took a while for Usuthu to get going, but they managed to carve out a chance in the 31st minute but George Chigova was on hand to make the save.
With AmaZulu struggling to carve out clear-cut chances, Boysen Mbatha tried his luck from distance in the 39th minute but his effort sailed over Chigova's crossbar.
Cavin Johnson's charges were the more dominant side in the encounter, but they were unable to find the equaliser in the opening 45 minutes.
AmaZulu eventually levelled in the 54th minute when Rhulani Manzini found the back of the net with a sublime free-kick from 30 yards out.
The hosts' joy was short-lived though, as Polokwane restored their lead two minutes later when Walter Musona fired home from 12 yards out after a goalkeeping error from Mbatha.
Usuthu continued to push for an equaliser, but it was Polokwane who got the next goal with Rodney Ramagalela firing past Mbatha at the near post.
Mhlengi Cele reduced the deficit once again in the 89th minute, but Rise and Shine managed to hold out to claim the vital win.