Expression and localization of VCX/Y proteins and their possible involvement in regulation of ribosome assembly during spermatogenesis

Variable Charge X/Y (VCX/Y) is a human testis-specific gene family that localized on X and Y chromosomes. In this study, VCY protein was expressed in E. coli in the form of glutathione-S-transferase (GST) fusion protein. With the purified fusion protein as antigen, the anti-GST-VCY antibody was generated and the localization of VCY protein in human testis was determined by immunohistochemistry. In the testis seminiferous epithelium. VCY proteins were highly expressed in nuclei of germ cells. Using propidium iodide staining and green fluorescent protein (GFP) tag technologies, VCY and VCX-8r proteins were mainly localized in the uucleoli of COST cells. In addition, the colocalization for VCY and VCX-8r in COS7 cells was also observed. With VCY cDNA as bait, a cDNA fragment of acidic ribosomal protein PO was obtained using yeast two-hybrid system. All the information above indicates that VCX/Y protein family might be involved in the regulation of ribosome assembly (hiring spermatogenesis.

Effects of Zn^（2＋） on root growth，cell division，and nucleoli of Allium cepa L．

The effects of different concentrations of Zn￣(2+)ion on root growth,cell division,and nucleoli of Allium cepa were studied. The test Zn￣(2+) ion concentration was made up from zinc sulphate (ZnSO4. 7H2O) ranging from 10￣(-7) to 10￣(-2) mol/L. The solutions were prepared in tap water (pH =6. 5).The results indicated that Zn￣(2+) could obviously inhibit root growth at concentrations from 10￣(-4)to 10￣(-2) mol/L.Roots treated with zinc sulphate showed the presence of c-mitosis, anaphase bridges,including sticky and fluidized bridges (at 10￣(-3) to 10￣(-2) mol/L) , chromosome stickiness, irregularly shaped nuclei, broken nuclei and micronuclei. A toxicity effect was also observed on the nucleoli using silver staining technique after 48h of treatment with 10￣(-4)to 10￣(-2) mol/L Zn￣(2+), e. g,the nucleolar particulate material scattered around the nucleoli in the nucleus of root tip cells.

Fluorescent Visualization of Nucleolar G-Quadruplex RNA and Dynamics of Cytoplasm and Intranuclear Viscosity

The nucleolus,the locus of ribosome biogenesis,was found to be the predominant intracellular target of a new fluorescent probe,V-P1.In solution,the probe demonstrated both a selectivity to RNA G-quadruplexes and a sensitivity to the viscosity,while G-quadruplex binding did not disturb the viscosity sensing.In cells,confocal and fluorescence lifetime imaging,combined with digestion and competition experiments,lent support to the hypothesis of an RNA-based G-quadruplex as the intracellular target,postulated to be nucleolar ribosomal RNA(rRNA).The probe demonstrated a high sensitivity to viscosity in both the cytoplasm and the nuclear compartment and was used to precisely interrogate the viscosity changes resulting from diverse stimuli,such as temperature,monensin treatment,and etoposide-induced apoptosis.Owing to the putative rRNA G-quadruplex binding in vitro and in vivo,and further combined with a relatively low degree of toxicity,the dye enabled the interrogation of cytoplasm and intranuclear viscosity changes under diverse conditions and found applications in studying the influence and significance of cytoplasm and intranuclear viscosity as well as in gaining insight into the native secondary structure of rRNA in nucleoli.