The Mkt1-G30D point mutation disrupts Mkt1/Pbp1 complex abundance and formation. (A) Table depicting Mkt1 aa polymorphisms in CEN.PK and S288C strains used in this study. (B) Western blot assessing Mkt1-Flag, Pab1, and Pbp1-HA protein levels in CEN.PK and S288C cells collected 3 h following switch from YPD to YPL medium. Note: Mkt1-Flag and Pbp1-HA levels in CEN.PK and S288C strains expressing Mkt1-D30 from the endogenous locus were diminished compared with strains expressing Mkt1-G30. (C) co-IP experiments testing interactions between Mkt1-Flag and Pbp1-HA in CEN.PK and S288C cells collected during log phase in YPD and 3 h after switching to YPL medium. Mkt1-Flag interacted poorly with Pbp1-HA in CEN.PK and S288C strains expressing Mkt1-D30 alleles. (D) Polysome profile of CEN.PK Mkt1-G30 cells collected 3 h following switch from YPD to YPL medium and associated western blot for Mkt1-Flag and Pbp1-HA in the collected fractions. Mkt1-G30-Flag and Pbp1-HA were detected in polysome fractions. (E) Polysome profile of CEN.PK Mkt1-D30 cells collected 3 h following switch from YPD to YPL medium and associated western blot for Mkt1-Flag and Pbp1-HA in the collected fractions. Mkt1-D30-Flag was less present in polysome fractions. Source data are available for this figure: SourceData F7.