1. Specialization and Targeting of B-Type Cyclins
Frederick R Cross, Maria Yuste-Rojas, Samantha Gray, Matthew D Jacobson 
Molecular Cell 
Volume 4, Issue 1, Pages (July 1999)
DOI: /S (00)

2. Figure 1
CLB5::CLB2 Inefficiently Rescues Replication Defects Due to the Absence of CLB5
Isogenic diploids of genotype clb3::LEU2/CLB3 clb4::HIS2/CLB4 clb6::ADE1/CLB6, and either clb5::ARG4/CLB5 or clb5::ARG4/CLB5::CLB2, were sporulated and tetrads dissected. Exponentially growing cultures of haploid segregants from these diploids were grown in YEPD at 30°C and analyzed by FACS analysis for DNA content. Representative strains of selected genotypes are presented.
Molecular Cell 1999 4, 11-19DOI: ( /S (00) )

3. Figure 2
Clb Protein and Kinase Levels for CLB5 Promoter-Driven Constructs
Exponentially growing cultures of cells with various HA-tagged coding sequences inserted precisely at the CLB5 locus were extracted and immunoprecipitated with anti-HA monoclonal antibody, followed by histone H1 kinase assay and Western blot for HA and for coimmunoprecipitated Cdc28p. (A) untagged (−), CLB5-HA, CLB5-Δdb-HA, CLB5-hpm-HA, and CLB2-HA. The apparently higher binding of Cdc28p to Clb5p-Δdb was not reproducibly observed (data not shown). (B) untagged (−), CLB5-HA (wt), CLB5-hpm-HA, and CLB5-KA,EA-HA; (C) untagged (−), CLB5-HA (wt), CLB5-hpm-HA, CLB5-KA,EA-HA, and CLB5-hpm,KA,EA-HA. An anti-Cdc28p Western blot was not performed in (C).
Molecular Cell 1999 4, 11-19DOI: ( /S (00) )

4. Figure 3
Cell Cycle Timing of Accumulation of Clb Protein and Associated Kinase Activity from CLB5-HA and CLB5::CLB2-HA
Exponentially growing cultures of cells with HA-tagged CLB5 and CLB2 coding sequences inserted precisely at the CLB5 locus were arrested with α factor and released. At the indicated time points, aliquots of the cultures were analyzed as in Figure 2A and Figure 2B for the level of immunoprecipitated HA-tagged protein and associated histone H1 kinase activity. The percentage of unbudded cells was determined, and FACS profiles were obtained for each time point (bottom).
Molecular Cell 1999 4, 11-19DOI: ( /S (00) )

5. Figure 4
Cell Cycle Timing of Accumulation of Clb Protein and Associated Kinase Activity from CLB5::CLB2-HA and CLB5::CLB2-Δdb-HA
Exponentially growing cultures of cells with HA-tagged CLB2 and CLB2-Δdb coding sequences inserted precisely at the CLB5 locus were analyzed as in Figure 3. The slight reduction in Clb2p-HA at the 60 min time point was not reproducible (Figure 3; data not shown).
Molecular Cell 1999 4, 11-19DOI: ( /S (00) )

6. Figure 5
The Effects of the Hydrophobic Patch Mutation and a Cyclin–Cdk Interface Mutation on CLB5 Replicative Function
FACS profiles were obtained from exponentially growing cultures in YEPD of a clb5::URA3 strain and isogenic derivatives in which clb5::URA3 was replaced with various CLB5 mutants, as indicated.
Molecular Cell 1999 4, 11-19DOI: ( /S (00) )

7. Figure 6
Lethality of GAL1::CLB5-Δdb: the KA,EA Mutation but Not the hpm Mutation Prevents Lethality of Overexpressed Stable Clb5
Wild-type strains were transformed with integrating plasmids containing GAL1::CLB5-Δdb containing either no additional mutations, the hpm mutation, the K253A,E282A mutation, or the combined hpm,KA,EA mutations. Stable transformants were identified and streaked on YEPD or YEPGal.
Molecular Cell 1999 4, 11-19DOI: ( /S (00) )

8. Figure 7 A Model for Modular Structure of Clb5
A spatial and functional distinction is proposed between the cyclin–Cdk interface (Jeffrey et al. 1995) partially inactivated by the KA,EA mutation, and the hydrophobic patch region (Schulman et al. 1998) mutated in the hpm mutation. The two mutations are proposed to interfere with Clb5p function in distinct ways, either lowering Clb5p-associated kinase, which is still targeted (KA,EA), or not affecting Clb5p-associated kinase levels but rather compromising targeting of the kinase (hpm).
Molecular Cell 1999 4, 11-19DOI: ( /S (00) )