Michael Matheson is being urged to consider his position after a Holyrood committee was told he described Chief Constable Phil Gormley's planned return to work as a "bad decision".
Andrew Flanagan was the chair of the Scottish Police Authority (SPA) when the decision was made to end the period of special leave for the top officer in Police Scotland - against whom several allegations of gross misconduct are being investigated.
Mr Gormley has been on leave since September, but at a meeting in November the SPA board agreed this could be brought to an end.
But Mr Flanagan said when he informed the Justice Secretary of this "he told me he thought it was a bad decision".
The former SPA chair stated: "It was clear to me that he did not want the Chief Constable to return at that point."
Mr Flanagan said after that he felt he had "no choice" but to reverse the decision, describing the Cabinet Secretary as being "very unhappy" about it.
Mr Matheson has previously insisted his concerns about the decision were related to the processes the SPA board followed, saying those investigating the allegations against Mr Gormley and other senior officers had not been informed of this the day before the Chief Constable was due to return to work.
But Mr Flanagan's evidence to MSPs on the Public Audit Committee led Conservative leader Ruth Davidson to say: "The Justice Secretary didn't merely ask questions - he leaned on the independent chair of an independent body, leaving him in no doubt that he should reverse his decision.
"This evidence given in the Scottish Parliament shows Michael Matheson was absolutely instrumental in preventing the chief constable from returning to work."
Labour justice spokesman Daniel Johnson also hit out, saying: "Mr Matheson misled Parliament, telling the chamber that he had only raised concerns about the decision-making process, not the decision to reinstate Phil Gormley itself.
"Furthermore, Andrew Flanagan revealed that he felt he had no choice but to reverse the decision following his meeting with the Justice Secretary, raising questions about whether Mr Matheson's actions where within the parameters of the law.
"It is clear that Michael Matheson has behaved inappropriately for a Cabinet Secretary, repeatedly obscuring the true details of exactly what happened in his meeting, or as it now has been revealed, meetings, with Andrew Flanagan, and he now must consider his position in the cabinet."
However Jenny Marra, the Public Audit Committee convener, said they had been "entirely underwhelmed by the accuracy and reliability of evidence" from former and current members of the SPA board.
And Nicola Sturgeon, who was asked about on the issue at First Minister's Questions, insisted the actions of his Justice Secretary had been "entirely appropriate".
The First Minister rejected claims Mr Matheson had "leaned on" the SPA, saying: "Andrew Flanagan...actually said at the committee this morning that he hadn't been requested by the Justice Secretary to change his decision, he had no option in his view but to change his decision because he couldn't answer the most basic questions about the process that had been followed."
Mr Gormley has been on special leave since September and denies the allegation against him.