From Polarity to Morphogenesis PAK Behaviors and Mechanism for Bud Sensing in Morphogenesis Checkpoint

Bud formation by Saccharomyces cerevisiae is a fundamental process for yeast proliferation. Bud emergence is initiated by the polarization of the cytoskeleton, leading to local secretory vesicle delivery and gulcan synthase activity. The master regulator of polarity establishment is a small Rho-family GTPase – Cdc42. Cdc42 forms a clustered patch at the incipient budding site in late G1 and mediates downstream events which lead to bud emergence. Cdc42 promotes morphogenesis via its various effectors. PAKs (p21-activated kinases) are important Cdc42 effectors which mediate actin cytoskeleton polarization and septin filament assembly. The PAKs Cla4 and Ste20 share common binding domains for GTP-Cdc42 and they are partially redundant in function. However, we found that Cla4 and Ste20 behaved differently during the polarization and this depended on their different membrane interaction domains. Also, Cla4 and Ste20 compete for a limited number of binding sites at the polarity patch during bud emergence. These results suggest that PAKs may be differentially regulated during polarity establishment.

Morphogenesis of yeast must be coordinated with the nuclear cycle to enable successful proliferation. Many environmental stresses temporarily disrupt bud formation, and in such circumstances, the morphogenesis checkpoint halts nuclear division until bud formation can resume. Bud emergence is essential for degradation of the mitotic inhibitor, Swe1. Swe1 is localized to the septin cytoskeleton at the bud neck by the Swe1-binding protein Hsl7. Neck localization of Swe1 is required for Swe1 degradation. Although septins form a ring at the presumptive bud site prior to bud emergence, Hsl7 is not recruited to the septins until after bud emergence, suggesting that septins and/or Hsl7 respond to a “bud sensor”. Here we show that recruitment of Hsl7 to the septin ring depends on a combination of two septin-binding kinases: Hsl1 and Elm1. We elucidate which domains of these kinases are needed, and show that artificial targeting of those domains suffices to recruit Hsl7 to septin rings even in unbudded cells. Moreover, recruitment of Elm1 is responsive to bud emergence. Our findings suggest that Elm1 plays a key role in sensing bud emergence.

U-Pb geochronology of the Santa Cruz Formation (early Miocene) at the Río Bote and Río Santa Cruz (southernmost Patagonia, Argentina): Implications for the correlation of fossil vertebrate localities

© 2016 Elsevier Ltd.The early Miocene Santa Cruz Formation (SCF) in southern Patagonia hosts the Santacrucian South American Land Mammal Age (SALMA), whose age is known mainly from exposures along the Atlantic coast. Zircon U-Pb ages were obtained from intercalated tuffs from four inland sections of the SCF: 17.36 ± 0.63 Ma for the westernmost Río Bote locality, and 17.04 ± 0.55 Ma-16.32 ± 0.62 Ma for central Río Santa Cruz localities. All ages agree with the bounding age of underlying marine units and with equivalent strata in coastal exposures. New ages and available sedimentation rates imply time spans for each section of ~18.2 to 17.36 Ma for Río Bote and 17.45-15.63 Ma for central Río Santa Cruz (Burdigalian). These estimates support the view that deposition of the SCF began at western localities ~1 Ma earlier than at eastern localities, and that the central Río Santa Cruz localities expose the youngest SCF in southern Santa Cruz Province. Associated vertebrate faunas are consistent with our geochronologic synthesis, showing older (Notohippidian) taxa in western localities and younger (Santacrucian) taxa in central localities. The Notohippidian fauna (19.0-18.0 Ma) of the western localities is synchronous with Pinturan faunas (19.0-18.0 Ma), but older than Santacrucian faunas of the Río Santa Cruz (17.2-15.6 Ma) and coastal localities (18.0-16.2 Ma). The Santacrucian faunas of the central Río Santa Cruz localities temporally overlap Colloncuran (15.7 Ma), Friasian (16.5 Ma), and eastern Santacrucian faunas.