Anti-CALML5 monoclonal antibody, clone 4G7.2

Human papillomavirus (HPV) types 16 and 18 are known to play a major role in cervical carcinogenesis. However, additional genetic alterations are required for the development and progression of cervical cancer. Our aim was to identify genes which are consistently down-regulated in cervical cancers (CxCa) and which are likely to contribute to malignant transformation. Microarray analyses of RNA from high-grade cervical precancers (CIN2/3) and CxCa were performed to screen for putative tumour suppressor genes (TSG) in predefined regions on chromosomes 4 and 10. Validation of the candidate genes was done by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) in 16 normal cervical tissues, 14 CIN2/3 and 16 CxCa. The two most promising genes, SORBS2 and CALML5, were expressed ectopically in various cell lines in order to analyse their functional activity. Reconstitution of SORBS2 expression resulted in a significant reduction in cell proliferation, colony formation and anchorage-independent growth in CaSki, HPKII and HaCaT cells, whereby anchorage-independent growth could only be investigated for CaSki cells. SORBS2 had no effect on cell migration. In contrast, reconstitution of CALML5 expression did not influence the phenotype of all cell lines tested. None of the genes could induce senescence or apoptosis. Our results underline a possible role of SORBS2 as a TSG in cervical carcinogenesis.

The structural component of wool and hair fibers, such as keratin-associated proteins (KAPs), has been well described, but the genetic determinants of fiber diameter are largely unknown. Here, we have used an iTRAQ-based proteomic approach to investigate differences in protein abundance among 18 samples from sheep and goats across a diverse range of fibers. We identified proteins with different abundance and are associated with variation in fiber features. Proteins with different abundance are mainly keratin or keratin-associated proteins (KRTAP11-1, KRT6A, KRT38), or are related to hair growth (DSC2, DSG3, EEF2, CALML5, TCHH, SELENBP1) and fatty acid synthesis (FABP4, FABP5). RNA-seq further confirmed the functional importance of the DSC2 gene in the determination of woolly phenotype in goat fibers. This comprehensive analysis of fibers from major fiber producing animals is the first to provide a list of candidate proteins that are involved in fiber formation. This list will be valuable asset for future studies into the molecular mechanisms that underlie fiber diversity. Biological significance: Proteins are the basis for animal-derived hair fibers, yet proteins conferring fiber structure and characteristics in sheep and goats are largely elusive. By examining 27 fibers samples representing 9 fiber types from sheep and goats through the iTRAQ approach, we show a list of differentially abundant proteins that are important to hair structural component, or genes related to hair growth and fatty acid synthesis. RNA-seq further validated the DSC2 gene is key to the woolly/straight hair phenotype in goats.