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A versatile toolbox for PCR‐based tagging of yeast genes: new fluorescent proteins, more markers and promoter substitution cassettes
Using the provided cassettes for N‐ and C‐terminal gene tagging or for deletion of any given gene, a set of only four primers is required, which makes this method very cost‐effective and reproducible. Expand
Evidence that the Ipl1-Sli15 (Aurora Kinase-INCENP) Complex Promotes Chromosome Bi-orientation by Altering Kinetochore-Spindle Pole Connections
IPL1 function in cells that cannot replicate their chromosomes but nevertheless duplicate their spindle pole bodies (SPBs) is investigated and the possibility that Ipl1-Sli15 facilitates bi-orientation by promoting turnover of kinetochore-SPB connections until traction of sister Kinetochores toward opposite spindle poles creates tension in the surrounding chromatin is raised. Expand
The Bub2p spindle checkpoint links nuclear migration with mitotic exit.
It is shown that Bfa1p and Bub2p bind the Ras-like GTPase Tem1p to the cytoplasmic face of the SPB that enters the bud, whereas the GDP/GTP exchange factor Lte1p is associated with the cortex of the bud. Expand
Modes of spindle pole body inheritance and segregation of the Bfa1p–Bub2p checkpoint protein complex
It is described that in unperturbed cells the ‘old’ SPB always migrates into the bud, and the Bfa1p localization is not determined by SPB inheritance, which may provide a mechanism to delay cell cycle progression when cytoplasmic microtubules fail to orient the spindle. Expand
Epitope tagging of yeast genes using a PCR‐based strategy: more tags and improved practical routines
In the yeast Saccharomyces cerevisiae, molecular biological techniques have been developed that use a simple PCR‐based strategy to introduce epitope tags to chromosomal loci to produce PCR amplificable modules. Expand
Nud1p links astral microtubule organization and the control of exit from mitosis
It is described that the core SPB component Nud1p is a key protein that functions in both budding yeast spindle pole body and mitotic exit, and the failure of Tem1p to interact with Cdc15p at the SPB probably prevents mitoticExit. Expand
Compartment‐specific aggregases direct distinct nuclear and cytoplasmic aggregate deposition
It is shown that JUNQ unexpectedly resides inside the nucleus, defining a new intranuclear quality control compartment, INQ, for the deposition of both nuclear and cytosolic misfolded proteins, irrespective of ubiquitination. Expand
The binding cascade of SecB to SecA to SecY E mediates preprotein targeting to the E. coli plasma membrane
The export of many E. coli proteins such as proOmpA requires the cytosolic chaperone SecB and the membrane-bound preprotein translocase, which has a dual function in stabilizing the precursor and in passing it on to membrane- bound SecA, the next step in the pathway. Expand
Δμ H+ and ATP function at different steps of the catalytic cycle of preprotein translocase
Preprotein translocation in E. coli requires ATP, the membrane electrochemical potential delta mu H+, and translocase, an enzyme with an ATPase domain (SecA) and the membrane-embedded SecY/E. StudiesExpand
Yeast Cdk1 translocates to the plus end of cytoplasmic microtubules to regulate bud cortex interactions
The Cdc28‐regulated localization of Kar9 is an integral part of the program that aligns spindles and is dependent upon the microtubule motor protein Kip2. Expand