League One promotion hopefuls Brentford and Leyton Orient both dropped points on Tuesday as Wolves increased their lead at the top.
Second-placed Brentford suffered a comprehensive 3-0 defeat at Rotherham United, but third-placed Orient failed to take advantage as they were held to a 1-1 home draw with Oldham Athletic.
Kieran Agard scored a first-half double to put Rotherham in command, the striker's first coming from the penalty spot as he took his tally for the season to 20 goals.
On loan Newcastle midfielder Haris Vuckic rounded off the win for Rotherham, who are now just four points adrift of third-placed Orient.
Meanwhile, a late Korey Smith equaliser denied Orient all three points against Oldham after defender Scott Cuthbert had given Russell Slade's side the lead.
And Wolves took full advantage of their rivals' slip-ups, as they moved four points clear at the summit after coming out on the right side of a six goal thriller with Colchester United, Kenny Jackett's men claiming a 4-2 win.
Goals from Alex Gilbey and David Wright gave Colchester hope and set up a tense finale after Wolves had surged ahead with three first-half goals from Michael Jacobs, David Edwards and James Henry.
But Nouha Dicko made sure of the points for the hosts in injury time.
At the other end of the table, Notts County moved off the bottom with a 3-1 win at Crewe Alexandra, Ronan Murray scoring a double to help them leapfrog Stevenage, who visit Coventry City on Wednesday.
Fellow strugglers Carlisle United and Shrewsbury Town played out a 0-0 draw, and Tranmere Rovers succumbed to a 2-1 home loss to Swindon Town.
Preston North End overcame Peterborough United 3-1 thanks to a late brace from Joe Garner, but MK Dons' play-off hopes were dealt a blow with a 1-0 reverse at home to Gillingham.
Bristol City were the big winners of the day, surging to a 5-0 victory over Port Vale - who had Carl Dickinson sent off when four goals down - thanks to doubles from Jay Emmanuel-Thomas and Sam Baldock.
Elsewhere, Conor Coady scored twice as Sheffield United beat Crawley Town 2-0, and Craig Westcarr netted both goals in Walsall's 2-0 triumph at Bradford City.