Beth Mooney‘s 61 not out steered the world T20 champions home with one ball to spare at Edgbaston chasing 154 to win.
After winning the sole Test match of the series, the visitors now lead 6-0 on points with just 10 left to play for, courtesy of two points for a win in the two remaining T20 clashes and three one day internationals.
Australia captain Alyssa Healy, who played despite managing two fractured fingers, won the toss and elected to field first.
Sophia Dunkley anchored the England innings as she hit 56 off 49 balls but wickets fell regularly at the other end.
Danni Wyatt, Alice Capsey and Nat Sciver-Brunt all departed for single figures.
But a middle order rally from captain Heather Knight (29) and Amy Jones, who was 40 not out, allowed England to post 153.
Healy fell early to the bowling of Lauren Bell, but a rapid 40 from Tahlia McGrath took the game away from England.
Ashleigh Gardner won the tourists the Test with the ball and took Australia close to the finishing line with 31 off 23 balls.
Two wickets in two balls from Sarah Glenn got rid of Gardner and Grace Harris.
But Mooney remained at the crease and hit nine fours in her 47-ball innings to make sure of victory.
England now likely need to win all five remaining matches in the series to reclaim the Ashes for the first time since 2014.
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