Structural and functional architecture of the yeast cell-cycle transcription factor swi6.
The structural and functional organisation of Swi6, a transcriptional regulator of the budding yeast cell cycle has been analysed by a combination of biochemical, biophysical and genetic methods. Limited proteolysis indicates the presence of a approximately 15 kDa N-terminal domain which is dispensable for Swi6 activity in vivo and which is separated from the rest of the molecule by an extended linker of at least 43 residues. Within the central region, a 141 residue segment that is capable of transcriptional activation encompasses a structural domain of approximately 85 residues. In turn, this is tightly associated with an adjacent 28 kDa domain containing at least four ankyrin-repeat (ANK) motifs. A second protease sensitive region connects the ANK domain to the remaining 30 kDa C-terminal portion of Swi6 which contains a second transcriptional activator and sequences required for heteromerisation with Swi4 or Mbp1. Transactivation by the activating regions of Swi6 is antagonised when either are combined with the central ankyrin repeat motifs. Hydrodynamic measurements indicate that an N-terminal 62 kDa fragment comprising the first three domains is monomeric in solution and exhibits an unusually high frictional coefficient consistent with the extended, multi-domain structure suggested by proteolytic analysis.
Division of Yeast Genetics, National Institute for Medical Research, London, UK.
S G
SG
Sedgwick
I A
IA
Taylor
A C
AC
Adam
A
A
Spanos
S
S
Howell
B A
BA
Morgan
M K
MK
Treiber
N
N
Kanuga
G R
GR
Banks
R
R
Foord
S J
SJ
Smerdon
D017487:Ankyrins / Q000737:chemistry; D002453:Cell Cycle / Q000502:physiology*; D002918:Chymotrypsin / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry* / Q000378:metabolism; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry; D011485:Protein Binding / Q000235:genetics; D011487:Protein Conformation; D012440:Saccharomyces / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins; D017421:Sequence Analysis; D017384:Sequence Deletion / Q000235:genetics; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism; D015533:Transcriptional Activation / Q000235:genetics; D014357:Trypsin / Q000378:metabolism; D014461:Ultracentrifugation
D017487:Ankyrins; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C083209:MBP1 protein, S cerevisiae; D010446:Peptide Fragments; C062214:SWI4 protein, S cerevisiae; C075111:SWI6 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D002918:Chymotrypsin; D014357:Trypsin