Constructed by a PCR-based method (Longtine et al., 1998).

Constructed by transforming BglII-digested YIp128-CDC3-mCherry (Gao et al., 2007) into YEF473A and integrating at the CDC3 locus.

Constructed by PCR-amplifying CHS2-GFP:HIS3MX6 and MYO2-GFP:HIS3 fragments from YEF5762 and YKT520, respectively, and transforming into YEF5804.

Constructed by transforming Y5005-8D with a MYO1-GFP:KanMX6 fragment that was PCR-amplified from YEF2293 (Luo et al., 2004), followed by integration of BglII-digested YIp128-CDC3-mCherry at the CDC3 locus.

Constructed by a PCR-based method(Longtine et al., 1998) using plasmids pFA6a-GFP-His3MX6 (Longtine et al., 1998) and pFA6a-GFP-URA3 (provided by C. Burd, University of Pennsylvania, Philadelphia, PA) as templates for tagging MYO1 and NUP57, respectively. The PCR fragments containing MYO1-GFP:His3MX6 and NUP57-GFP:URA3 were transformed sequentially into strain RLY1 (Winter et al., 1999).

Constructed by transforming PCR-amplified NUP57-GFP:URA3 from YEF6113 into strains XDY41, YJL488A, YJL335A, YJL222A, and XDY254, respectively.

Constructed by transforming PCR-amplified BNI5-mCherry:KanMX6 from XDY211 into XDY41.

Constructed by transforming PCR-amplified bni5Δ::His3MX6 from XDY254 into YJL448A and XDY289, respectively.

Strain XDY218 was first transformed by BglII-digested YIp128-CDC3-mCherry and then streaked on a SC-Leu + 5-FOA plate to select for the loss of plasmid pRS316-MYO1-TD2-GFP. The resulting strain was then transformed with plasmid pRS316-MYO1-C-GFP.

Constructed by streaking XDY211 on a SC + 5-FOA plate to select for the loss of plasmid pRS316-N-NotI-MYO1.

Strain RNY1264 (iqg1Δ::His3MX6 MYO1-GFP:KanMX6 [YCp50-IQG1]) was transformed with pRS315-IQG1-GST or pRS315-IQG1(Δ2-411)-GST, and the resulting strains were streaked on a SC-Leu + 5-FOA plate to eliminate YCp50-IQG1.

Constructed by transforming BglII-digested YIp211-CDC3-mCherry into strains RNY2594 and RNY2595 and integrating at the CDC3 locus.

Constructed by transforming the plasmid into YEF473A and Y5005-8D, respectively.

Strains carrying different myo1 alleles were transformed with plasmid pRS316-MYO1. The resulting strains were either 3HA-tagged or deleted for BNI5 using a PCR-based method (Longtine et al., 1998), and then selected for the loss of the MYO1 plasmid by growing cells in non-selective YPD medium.

Constructed by transforming a BNI5-mCherry:KanMX6 fragment that was PCR-amplified from plasmid pFA6a-mCherry-KanMX6 into YJL489A carrying plasmid pRS316-N-NotI-MYO1 (Caviston et al., 2003).

Strain RNY970 (shown in the table) was streaked from a SC-His plate onto a SC-His + 5-FOA plate; the resulting single colonies were checked by PCR to confirm the loss of the IQG1 plasmid. The resulting iqg1Δ::His3MX6 strain was then transformed with indicated plasmids to generate strains XDY218, XDY295, XDY296, and XDY300.

Strains XDY41, XDY287, YJL335A, YJY222A, and YJY488A were transformed with plasmid pRS316-MYO1. The resulting strains were then transformed with BglII-digested integrative plasmid YIp128-CDC3-mCherry and transformants were selected on SC-Leu plates at 25°C and then grown in non-selective YPD medium to lose the MYO1 plasmid.