The two-time Rugby World Cup winner, who played 16 Tests for the Black Ferns, took up refereeing in 2015.
She has subsequently controlled games in New Zealand's Mitre 10 Cup, including a Ranfurly Shield challenge, and the Heartland competition.
Her goal was always to referee during the Women's World Cup being staged in New Zealand in 2021 and the Six Nations will be an important step towards achieving that.
"For me the women's World Cup would be amazing because I've played in a couple of them and it's in New Zealand, so that's a massive goal of mine," she told rwcwomens.com.
Mahoney will control the Ireland v Italy game in Dublin on March 8 and a week later will be on Cardiff Arms Park for the Wales v Scotland game.
World Rugby chairman Sir Bill Beaumont said: "The Women's Six Nations Championship will kick-start a new decade that we hope will see continued and unprecedented growth in women's rugby.
"With our strong focus on developing women in rugby across the game, it is very pleasing that we have been able to appoint female only teams of three on-field match officials for all 15 of the Women's Six Nations games.
"The match officials are continuing their preparations for Rugby World Cup 2021 and I'm confident this squad has got an excellent balance of youth and experience to deliver a very successful tournament," he said.
World Rugby 15s high performance match official manager Alain Rolland congratulated Mahoney, and Australian Amber McLachlan, who will also be making her debut.
"There has been incredible work done bringing this team of match officials to the best possible level and we believe they are in very good shape less than two years away from New Zealand 2021.
"They are a dedicated group with high standards, and we are excited to see new referees coming from our high-performance programmes and other women's competitions making it to Test level.
"Our goal is always clear, consistent and accurate decision-making and while there is no change in emphasis, we will continue to work closely with teams to ensure the best possible experience for players and fans," Rolland said.