Tag Archives: congratulations

Congratulations to Yansong Miao on his new paper!

Yansong Miao‘s new paper is out now at The Proceedings of the National Academy of Sciences.  Congratulations to Yansong on his great work!  The abstract is below.  The PDF can be downloaded from PNAS here.

Cell-cycle regulation of formin-mediated actin cable assembly. Miao Y, Wong CC, Mennella V, Michelot A, Agard DA, Holt LJ, Yates JR 3rd, Drubin DG. Proc Natl Acad Sci U S A. 2013 Nov 19;110(47):E4446-E4455. Epub 2013 Oct 16.  PMID: 24133141

Abstract

Assembly of appropriately oriented actin cables nucleated by formin proteins is necessary for many biological processes in diverse eukaryotes. However, compared with knowledge of how nucleation of dendritic actin filament arrays by the actin-related protein-2/3 complex is regulated, the in vivo regulatory mechanisms for actin cable formation are less clear. To gain insights into mechanisms for regulating actin cable assembly, we reconstituted the assembly process in vitro by introducing microspheres functionalized with the C terminus of the budding yeast formin Bni1 into extracts prepared from yeast cells at different cell-cycle stages. EM studies showed that unbranched actin filament bundles were reconstituted successfully in the yeast extracts. Only extracts enriched in the mitotic cyclin Clb2 were competent for actin cable assembly, and cyclin-dependent kinase 1 activity was indispensible. Cyclin-dependent kinase 1 activity also was found to regulate cable assembly in vivo. Here we present evidence that formin cell-cycle regulation is conserved in vertebrates. The use of the cable-reconstitution system to test roles for the key actin-binding proteins tropomyosin, capping protein, and cofilin provided important insights into assembly regulation. Furthermore, using mass spectrometry, we identified components of the actin cables formed in yeast extracts, providing the basis for comprehensive understanding of cable assembly and regulation.

Miao 2013 PNAS Figure 2

Average intensity projections along the z-axis of WT cells stained with Alexa-568 phalloidin to label actin filaments and expressing GFP-Tub1 as a cell-cycle stage indicator, obtained using 3D SIM microscopy.

Congratulations to Yidi Sun on her new paper!

Yidi Sun‘s new paper is out now as an electronic publication ahead of print in the Journal of Cell Science.  Congratulations to Yidi on her great work!  The abstract is below.  The PDF can be downloaded from JCS here.

The functions of anionic phospholipids during clathrin-mediated endocytosis site initiation and vesicle formation. Sun Y, Drubin DG. J Cell Sci. 2012 Oct 24. PMID: 23097040.

Abstract

Anionic phospholipids PI(4,5)P(2) and phosphatidylserine (PS) are enriched in the cytosolic leaflet of the plasma membrane where endocytic sites form. In this study, we investigated the roles of PI(4,5)P(2) and PS in clathrin-mediated endocytosis (CME) site initiation and vesicle formation in Saccharomyces cerevisiae. Live-cell imaging of endocytic protein dynamics in an mss4(ts) mutant, which has severely reduced PI(4,5)P(2) levels, revealed that PI(4,5)P(2) is required for endocytic membrane invagination but is less important for endocytic site initiation. We also demonstrated that in various deletion mutants of genes encoding components of the Rcy1-Ypt31/32 GTPase pathway, endocytic proteins dynamically assemble not only on the plasma membrane but also on intracellular membrane compartments, which are likely derived from early endosomes. In rcy1Δ cells, fluorescent biosensors indicated that PI(4,5)P(2) only localized to the plasma membrane while PS localized to both the plasma membrane and intracellular membranes. Furthermore, we found that polarized endocytic patch establishment is defective in the PS-deficient cho1Δ mutant. We propose that PS is important for directing endocytic proteins to the plasma membrane and that PI(4,5)P(2) is required to facilitate endocytic membrane invagination.

Congratulations to Yidi Sun on her new paper!

Yidi Sun‘s new paper is out now as an electronic publication ahead of print in the journal Molecular Biology of the Cell.  Congratulations to Yidi on her great work!  The abstract is below.  The PDF can be downloaded from MBoC here.

Orm protein phosphoregulation mediates transient sphingolipid biosynthesis response to heat stress via the Pkh-Ypk and Cdc55-PP2A pathways. Sun Y, Miao Y, Yamane Y, Zhang C, Shokat KM, Takematsu H, Kozutsumi Y, Drubin DG. Mol Biol Cell. 2012 Apr 25. PMID: 22535525.

Abstract

Sphingoid intermediates accumulate in response to a variety of stresses, including heat, and trigger cellular responses. However, the mechanism by which stress affects sphingolipid biosynthesis has yet to be identified. Recent studies in yeast suggested that sphingolipid biosynthesis is regulated through phosphorylation of the Orm proteins, which in humans are potential risk factors for childhood asthma. Here, we demonstrate that Orm phosphorylation status is highly responsive to sphingoid bases. We also demonstrate by monitoring temporal changes in Orm phosphorylation and sphingoid base production in cells inhibited for Ypk1 protein kinase activity, that Ypk1 transmits heat stress signals to the sphingolipid biosynthesis pathway via Orm phosphorylation. Our data indicate that heat-induced sphingolipid biosynthesis in turn triggers Orm protein dephosphorylation, making the induction transient. We identified Cdc55-PP2A (protein phosphatase 2A) as a key phosphatase that counteracts Ypk1 activity in Orm mediated sphingolipid biosynthesis regulation. In total, our study reveals a mechanism through which the conserved Pkh-Ypk kinase cascade and Cdc55-PP2A facilitate rapid, transient sphingolipid production in response to heat stress through Orm protein phosphoregulation. We propose that this mechanism serves as the basis for how Orm phosphoregulation controls sphingolipid biosynthesis in response to stress in a kinetically coupled manner.

Rebecca Lu is qualified!

The Drubin/Barnes Lab would like to congratulate Rebecca Lu who passed her qualifying exam on Wednesday, April 18!

Here we recount the epic saga of Rebecca’s Ph.D qualification:

After she wrote her proposal, her chair asked her to rewrite it.  Rebecca is smarter than Randy Schekman though.  She just made a copy.

Rebecca brought food for her committee, and Doug Koshland said, ‘I’m gonna have a donut.  Does anyone want one?’ That’s when Rebecca realized that a Qualifying Exam is like some weird quiz where your committee reveals the answers before asking their questions.

During Rebecca’s exam, her committee didn’t ask her a question for 30 minutes.  That would have been a REALLY long question, after all.

Rebecca closed her eyes a lot during her Qual.  She wasn’t sleeping.  She just drew a picture of Randy handing her the Nobel Prize on the back of her eyelids.

You can’t please all the people all the time.  Fortunately, all those people weren’t at Rebecca’s Qualifying Exam.

When her exam was over, Randy said, “Guess what.  You passed.”  Rebecca told Randy, “Dude, you gotta give me time to guess.  If you’re gonna quiz me, you must insert a pause in there.”

Akemi Kunibe was the last grad student in the Drubin/Barnes Lab to pass her Qual.  She’s smart.  She was a tough act to follow.  Rebecca will be an especially hard act to follow, cuz when she finished her Qual, she took all of the dry-erase markers with her.

Rebecca doesn’t know the meaning of the word “fail.”  And that is kinda worse than not passing in a way, if you think about it.  She’s a Ph.D candidate, but she still doesn’t understand simple words.

Congratulations Rebecca Lu!  And thank you Mitch Hedberg for the jokes!