Cartier departed with five ships. Jean-François de La Rocque de Roberval was appointed as Lieutenant General, in overall command of the expedition, but Cartier sailed ahead.

Cartier established a settlement called Charlesbourg-Royal near Stadacona (Quebec City). He built a fort and began preparing for the winter. Relations with the local Iroquoians deteriorated. It is possible Donnacona had died in France, and the local tribes were no longer as trusting.

Cartier discovered what he believed were diamonds and gold at a location he named Cap Diamant (Cape Diamond). These 'diamonds' were later found to be quartz crystals, and the 'gold' was pyrite ('fool's gold').

Cartier, convinced of the value of his discoveries, abandoned the colony (likely due to the increased hostility of the Iroquois and the harsh winter) and sailed back to France before Roberval arrived with the main body of colonists and supplies.

Cartier encountered Roberval at St. John's, Newfoundland. Roberval ordered him to return to Charlesbourg-Royal, but Cartier disobeyed and continued to France under the cover of darkness.

Roberval continued to Charlesbourg-Royal and attempted to establish a colony, but it was ultimately unsuccessful due to the harsh conditions, disease, and conflict with the Iroquoians. The survivors returned to France in 1543.