In silico studies of potential phosphoresidues in the human nucleophosmin/B23: its kinases and related biological processes

Human nucleophosmin/B23 is a phosphoprotein involved in ribosome biogenesis, centrosome duplication, cancer, and apoptosis. Its function, localization, and mobility within cells, are highly regulated by phosphorylation events. Up to 21 phosphosites of B23 have been experimentally verified even though the corresponding kinase is known only for seven of them. In this work, we predict the phosphorylation sites in human B23 using six kinase-specific servers (KinasePhos 2.0, PredPhospho, NetPhosK 1.0, PKC Scan, pkaPS, and MetaPredPS) plus DISPHOS 1.3, which is not kinase specific. The results were integrated with information regarding 3D structure and residue conservation of B23, as well as cellular localizations, cellular processes, signaling pathways and protein-protein interaction networks involving both B23 and each predicted kinase. Thus, all 40 potential phosphosites of B23 were predicted with significant score (>0.50) as substrates of at least one of 38 kinases. Thirteen of these residues are newly proposed showing high susceptibility of phosphorylation considering their solvent accessibility. Our results also suggest that the enzymes CDKs, PKC, CK2, PLK1, and PKA could phosphorylate B23 at higher number of sites than those previously reported. Furthermore, PDK, GSK3, ATM, MAPK, PKB, and CHK1 could mediate multisite phosphorylation of B23, although they have not been verified as kinases for this protein. Finally, we suggest that B23 phosphorylation is related to cellular processes such as apoptosis, cell survival, cell proliferation, and response to DNA damage stimulus, in which these kinases are involved. These predictions could contribute to a better understanding, as well as addressing further experimental studies, of B23 phosphorylation.     

FLRT2 promotes cellular proliferation and inhibits cell adhesion during chondrogenesis

One of the earliest events during chondrogenesis is the formation of condensations, a necessary pre‐requisite for subsequent differentiation of a chondrogenic phenotype. Members of the Fibronectin Lecucine Rich Transmembrane (FLRT) proteins have been shown to be involved in cell sorting and neurite outgrowth. Additionally, FLRT2 is highly expressed at putative sites of chondrogenic differentiation during craniofacial development. In this study, we demonstrate that FLRT2 plays a role in mediating cell proliferation and cell–cell interactions during early chondrogenesis. Clones of stable transfectants of a murine chondroprogenitor cell line, ATDC5, were established in which FLRT2 was knocked down or overexpressed. Cells in which FLRT2 was knocked down proliferated at a slower rate compared to control wild‐type ATDC5 cells or those containing a non‐coding shRNA. In addition, FLRT2 knockdown cells formed numerous lectin peanut agglutinin (PNA) stained aggregates and exhibited higher expression of the cell adhesion molecule, N‐cadherin. In an in vitro wound healing assay, fewer FLRT2 knockdown cells appeared to migrate into the defect. Surprisingly, the FLRT2 knockdown cells demonstrated increased formation of Alcian blue‐stainable extracellular matrix, suggesting that their reduced aggregate formation did not inhibit subsequent chondrogenic differentiation. The opposite trends were observed in ATDC5 clones that overexpressed FLRT2. Specifically, FLRT overexpressing cells proliferated faster, formed fewer PNA‐positive aggregates, accumulated increased Alcian blue‐positive matrix, and migrated faster to close a wound. Collectively, our findings provide evidence for a role of FLRT2 in enhancing cell proliferation and reducing intercellular adhesion during the early stages of chondrogenesis. J. Cell. Biochem. 112: 3440–3448, 2011. © 2011 Wiley Periodicals, Inc.     

Saitohin,which is nested within the tau gene,interacts with tau and Abl and its human‐specific allele influences Abl phosphorylation

Saitohin (STH) is a gene unique to humans and their closest relatives whose function is not yet known. STH contains a single polymorphism (Q7R); the Q allele is human‐specific and confers susceptibility to several neurodegenerative diseases. In previous work, we discovered that STH interacts with Peroxiredoxin 6 (Prdx6), a unique member of that family which is bifunctional and whose levels increase in Pick's disease. In this study, we report that STH also interacts with tau and the non‐receptor tyrosine kinase c‐Abl (Abl). Furthermore, Abl phosphorylates STH on its single tyrosine residue and STH increases tyrosine phosphorylation by Abl. The effect of Saitohin on Abl‐mediated phosphorylation appears to be allele‐specific, providing evidence for a new cellular function for STH. J. Cell. Biochem. 112: 3482–3488, 2011. © 2011 Wiley Periodicals, Inc.     

A preliminary assessment of the impact of alien rainbow trout (Oncorhynchus mykiss) on indigenous fishes of the upper Berg River,Western Cape Province,South Africa

Impacts of alien rainbow trout (Oncorhynchus mykiss) on critically endangered Berg River redfin (Pseudobarbus burgi), Cape kurper (Sandelia capensis) and Cape galaxias (Galaxias zebratus) in the upper Berg River were investigated in terms of predation and spatial interactions. Trout stomach contents revealed that invertebrates dominate trout diet within the study area, whilst only six fish were recovered from 45 stomachs. The apparent low fish predation success of O. mykiss within the stream suggests a smaller impact compared to that of other alien piscivores such as bass (Micropterus spp.). Galaxias zebratus was the only fish species identified as prey, and its conservation status in the river requires further investigation. Snorkelling surveys revealed that rainbow trout co-exist with S. capensis and adult P. burgi within pools on this river. Galaxias zebratus was absent from the pools, while P. burgi juveniles were segregated from rainbow trout along a depth gradient, possibly indicating avoidance behaviour. Sandelia capensis juveniles may avoid predation by hiding under rocks. Rainbow trout probably compete with indigenous fish for food and space in the pools, though this could not be quantified. The impacts of O. mykiss on all indigenous fauna within the river are likely to be density-dependent.