RESUMO
We identified 7 SHP-1 (PTPN6) transcripts using epithelial cancer-derived cell lines. Four were shown to utilize the epithelial promoter 1 to transcribe a full-length, a partial (exon 3) or complete (exons 3 & 4) deletion of the N-SH2 domain, and also a non-coding transcript having a stop codon caused by a frame shift due to intron 2 retention. Three additional transcripts were shown to utilize the hematopoietic promoter 2 to transcribe a full-length, a partial (exon 3) deletion of the N-SH2 domain and a non-coding transcript with intron 2 retention. We show that endogenous proteins corresponding to the open-reading-frame (ORF) transcripts are produced. Using GST-fusion proteins we show that each product of the ORF SHP-1 transcripts has phosphatase activity and isoforms with an N-SH2 deletion have increased phosphatase activity and novel protein-protein interactions. This study is the first to document utilization of promoter 2 by SHP-1 transcripts and a noncoding transcript in human epithelial cells.
Assuntos
Neoplasias Epiteliais e Glandulares/genética , Neoplasias Epiteliais e Glandulares/metabolismo , Proteína Tirosina Fosfatase não Receptora Tipo 6/genética , Proteína Tirosina Fosfatase não Receptora Tipo 6/metabolismo , Processamento Alternativo , Linhagem Celular Tumoral , Éxons , Mutação da Fase de Leitura , Células HEK293 , Humanos , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/metabolismo , Células Jurkat , Células MCF-7 , Regiões Promotoras Genéticas , Ligação Proteica , Proteína Tirosina Fosfatase não Receptora Tipo 6/química , RNA não Traduzido/genética , RNA não Traduzido/metabolismoRESUMO
A yield enhancement technology for use in influenza vaccine manufacturing has been developed to maximize the recovery of influenza virus from allantoic fluid of virus-infected chick embryos; the standard raw material for influenza vaccine. Virus associated with amorphous debris in the allantoic fluid can be dissociated from the debris and recovered, thereby increasing viral yield. Dissociation can be achieved by subjecting the virus-debris complex to conditions of increased ionic strength at defined pH. Multifold increases in viral yield per ml of allantoic fluid were observed. The degree of yield enhancement is strain-specific, however, increases were observed in all type A and type B influenza strains tested. The heightened influenza virus recoveries can facilitate rapid vaccine manufacture, with increased numbers of doses produced, and may become essential at a time of influenza pandemic.