Cloning and expression of the essential gene for poly(A) polymerase from S. cerevisiae.

Poly(A) polymerase is essential for the maturation of messenger RNA, adding tracts of adenosine residues to the 3' end of precursor RNA generated by endonucleolytic cleavage. This mechanism of mRNA 3' processing seems to be similar in yeast and in higher eucaryotes, although there are differences in the recognition signals in the pre-mRNA. Here we describe the cloning of the gene for yeast poly(A) polymerase. The enzyme is encoded by a single and essential gene located near the centromere on the left arm of chromosome 11. Poly(A) polymerase purified from recombinant Escherichia coli has the same physical and biochemical properties as the yeast enzyme. The yeast poly(A) polymerase shares features of sequence with its mammalian homologue.     

A recycling pathway between the endoplasmic reticulum and the Golgi apparatus for retention of unassembled MHC class I molecules

Assembly of class I major histocompatibility complex (MHC) molecules involves the interaction of two distinct polypeptides (the heavy and light chains) with peptide antigen. Cell lines synthesizing both chains but expressing low levels of MHC class I molecules on their surface as a result of a failure in assembly and transport have been identified. We now report that although the apparent steady-state distribution in these cells of class I molecules is in the endoplasmic reticulum (ER), the molecules in fact are recycled between the ER and Golgi, rather than retained in the ER. This explains the failure of class I molecules to negotiate the secretory pathway. Class I molecules do not seem to be modified by Golgi enzymes, suggesting that the proteins do not reach the Golgi apparatus during recycling. But morphological and subcellular fractionation evidence indicates that they pass through the cis Golgi or a Golgi-associated organelle, which we postulate to be the recycling organelle. This compartment, which we call the 'cis-Golgi network', would thereby be a sorting organelle that selects proteins for return to the ER.     

Countercurrent chromatography meets MS

Interfacing countercurrent chromatography with thermospray mass spectrometry provides a new analytical tool for detecting nonvolatile, hydrophilic or thermally unstable bioactive natural products.     

A superantigen encoded in the open reading frame of the 3' long terminal repeat of mouse mammary tumour virus.

Mice express a collection of superantigens, which bind to class II major histocompatibility proteins and interact with T cells bearing particular V beta chains as part of their alpha beta receptors. These superantigens have been suggested to be encoded by exogenous or endogenous mouse mammary tumour viruses. One such superantigen is now shown to be encoded in the open reading frame of the long terminal repeat of a mammary tumour virus, a gene of previously unknown function.     

Comparison of counterimmunoelectrophoresis (CIE), latex agglutination (LA) and staphylococcal coagglutination (COAG) in pneumococcal antigen detection in vitro

CIE was compared to agglutination assays employing commercial kits (Directigen, Phadebact), as well as our own LA and COAG reagents, in detection of pneumococcal capsular polysaccharide (PCP) antigens in vitro. Directigen provided the most sensitive assay. CIE was of comparable sensitivity except for PCP antigen types 7 and 14.     

Androstenedione or corticosterone treatment during pregnancy alters estrous cycle of adult female offspring in mice

Female offspring from mice injected with androstenedione during late pregnancy showed lengthened vaginal cycles, persistent estrus and decreased incidence of pro-estrus and diestrus, whilst offspring from mice injected with corticosterone showed increased incidence of diestrus. These observations give qualified support to the hypothesis that stress during pregnancy alters the female offspring reproductive system through the action of adrenal steroids.     

Abnormal development of phialides in a strain ofAspergillus niger

Summary AnAspergillus niger mutant strain (hpp) produces an average of 4.1% of conidiophores with phialide proliferations. Increased frequency of proliferations could be induced on all studied strains by growth on potato dextrose agar. The characteristic is recessive and seems to be due to a pleiotropic effect of the mutation for olive conidia color.Acknowledgments. The authors are indebted to CNPq for financial assistance provided with grant PIG/SIP 04/053 as well as the scholarships Pesquisador Científico (R.B.Jr) and Iniciação Científica and Aperfeiçoamento (G.U.V.).     

Morphological transformation in vivo of human uterine cervix with papillomavirus from condylomata acuminata

Carcinoma of the human uterine cervix has been associated with several infectious agents including papillomavirus. Papillomavirus group-specific antigen (GSA) and viral particles have been demonstrated in human condylomata acuminata (CA) and flat warts of the uterine cervix. Cell alterations consisting of nuclear enlargement, hyperchromasia, irregularity, binucleation and cytoplasmic clearing (koilocytosis) are often interpreted as mild to moderate dysplasia. Present evidence that human papillomavirus (HPV) is responsible for the development of these lesions relies on the association of GSA and virus particles in the affected tissue, fulfilling the first two of Koch's postulates. Direct proof of an aetiological relationship, however, requires induction of the CA change in normal, human uterine cervix after exposure to papillomavirus. Infecting human subjects with HPV is ethically unacceptable and no satisfactory alternative systems have been defined. Also, human cell cultures do not support growth or transformation by HPV. Here we report the first demonstration of the morphological transformation of human tissues with a human papillomavirus under controlled, experimental conditions. 'Transformation' is used here in its literal sense to refer to a heritable morphological alteration in the appearance of the cells. The use of this term does not indicate that the changes described are neoplastic, but they are identical to the dysplastic changes found in biopsies of uterine cervical CA. Our results demonstrate the direct involvement of CA virus in dysplastic change of human cervical tissue and indicate that the experimental system described may be useful in elucidating the contribution of human papillomaviruses to the pathogenesis of human cervical cancer.