Leyton Orient began Tuesday one point ahead of Kenny Jackett's side, but slipped up on the road.
And that allowed Wolves to go top as they extended their unbeaten league record to 11 matches.
Leigh Griffiths opened the scoring at Molineux after 17 minutes and Wolves, who dominated proceedings, doubled their lead when David Edwards netted just before half-time.
That was enough for the hosts as Tranmere came crashing back to earth after Saturday's 5-1 win at Coventry City.
Bristol City might be in the relegation zone but they showed enough fight to come from behind twice in a 2-2 home draw against Orient.
A 13th league goal of the season for Republic of Ireland forward David Mooney gave Orient the lead in the 23rd minute.
Sam Baldock responded after the break for City and, although Dean Cox restored Orient's advantage, a Scott Cuthbert own goal ensured the sides shared the spoils.
The pressure on Peterborough United boss Darren Ferguson will continue to grow after their winless run extended to six league matches.
Brentford condemned Peterborough to their fifth loss in a row, as goals in the last 10 minutes from George Saville and Clayton Donaldson gave them a 3-2 home win.
Peterborough now sit fifth in the table, with Brentford and Preston North End - who won 2-0 at Port Vale - moving above them.
Joe Garner scored twice for Preston, who won despite having Neil Kilkenny sent off after just 13 minutes.
At the wrong end of the table, rock-bottom Notts County ended a run of three league losses with a point in a 1-1 draw at Bradford City.
The 23rd-placed Crewe Alexandra lost for the third league match in a row as they were beaten 2-1 at Carlisle United, while Sheffield United are still in the relegation zone despite a 1-1 draw against Walsall.
Other results saw Gillingham edge Stevenage 3-2, while Colchester United notched a 3-1 home win over Milton Keynes Dons.
Elsewhere, Rotherham United strolled to a 3-0 success against Coventry City, Oldham Athletic won 2-1 at Shrewsbury Town and Crawley Town and Swindon Town played out a goalless draw.
