RESUMO
The prostate-specific antigen-related serine protease gene, kallikrein 4 (KLK4), is expressed in the prostate and, more importantly, overexpressed in prostate cancer. Several KLK4 mRNA splice variants have been reported, but it is still not clear which of these is most relevant to prostate cancer. Here we report that, in addition to the full-length KLK4 (KLK4-254) transcript, the exon 1 deleted KLK4 transcripts, in particular, the 5'-truncated KLK4-205 transcript, is expressed in prostate cancer. Using V5/His6 and green fluorescent protein (GFP) carboxy terminal tagged expression constructs and immunocytochemical approaches, we found that hK4-254 is cytoplasmically localized, while the N-terminal truncated hK4-205 is in the nucleus of transfected PC-3 prostate cancer cells. At the protein level, using anti-hK4 peptide antibodies specific to different regions of hK4-254 (N-terminal and C-terminal), we also demonstrated that endogenous hK4-254 (detected with the N-terminal antibody) is more intensely stained in malignant cells than in benign prostate cells, and is secreted into seminal fluid. In contrast, for the endogenous nuclear-localized N-terminal truncated hK4-205 form, there was less difference in staining intensity between benign and cancer glands. Thus, KLK4-254/hK4-254 may have utility as an immunohistochemical marker for prostate cancer. Our studies also indicate that the expression levels of the truncated KLK4 transcripts, but not KLK4-254, are regulated by androgens in LNCaP cells. Thus, these data demonstrate that there are two major isoforms of hK4 (KLK4-254/hK4-254 and KLK4-205/hK4-205) expressed in prostate cancer with different regulatory and expression profiles that imply both secreted and novel nuclear roles.
Assuntos
Núcleo Celular/química , Citoplasma/química , Calicreínas/análise , Neoplasias da Próstata/metabolismo , Androgênios/metabolismo , Linhagem Celular Tumoral , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Glicosilação , Humanos , Calicreínas/genética , Calicreínas/metabolismo , Masculino , Neoplasias da Próstata/química , Neoplasias da Próstata/genética , Biossíntese de Proteínas , Isoformas de Proteínas/análise , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Sêmen/metabolismoRESUMO
In a recent case of scrub typhus in Australia, Orientia tsutsugamushi isolated from the patient's blood was tested by sequence analysis of the 16S rDNA gene. The sequence showed a strain of O. tsutsugamushi that was quite different from the classic Karp, Kato, and Gilliam strains. The new strain has been designated Litchfield.
Assuntos
Orientia tsutsugamushi/classificação , Adulto , Animais , Austrália , DNA Ribossômico/análise , Feminino , Humanos , Masculino , Camundongos , Orientia tsutsugamushi/genética , RNA Ribossômico 16S/análise , Tifo por Ácaros/microbiologiaRESUMO
The ITS sequences of Acropora spp. are the shortest so far identified in any metazoan and are among the shortest seen in eukaryotes; ITS1 was 70-80 bases, and ITS2 was 100-112 bases. The ITS sequences were also highly variable, but base composition and secondary structure prediction indicate that divergent sequence variants are unlikely to be pseudogenes. The pattern of variation was unusual in several other respects: (1) two distinct ITS2 types were detected in both A. hyacinthus and A. cytherea, species known to hybridize in vitro with high success rates, and a putative intermediate ITS2 form was also detected in A. cytherea; (2) A. valida was found to contain highly (29%) diverged ITS1 variants; and (3) A. longicyathus contained two distinct 5.8S rDNA types. These data are consistent with a reticulate evolutionary history for the genus Acropora.
Assuntos
Cnidários/genética , DNA Ribossômico/genética , Evolução Molecular , Animais , Sequência de Bases , Cnidários/classificação , Primers do DNA/genética , DNA Ribossômico/química , Variação Genética , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 5,8S/genética , Especificidade da EspécieRESUMO
Since both internal (class-level) and external relationships of the Cnidaria remain unclear on the basis of analyses of 18S and (partial) 16S rDNA sequence data, we examined the informativeness of the 5'-end of the 23S-like rDNA. Here we describe analyses of both primary and predicted secondary structure data for this region from the ctenophore Bolinopsis sp., the placozoan Trichoplax adhaerens, the sponge Hymeniacidon heliophila, and representatives of all four cnidarian classes. Primary sequence analyses clearly resolved the Cnidaria from other lower Metazoa, supported sister group relationships between the Scyphozoa and Cubozoa and between the Ctenophora and the Placozoa, and confirmed the basal status of the Anthozoa within the Cnidaria. Additionally, in the ctenophore, placozoan and sponge, non-canonical base pairing is required to maintain the secondary structure of the B12 region, whereas amongst the Cnidaria this is not the case. Although the phylogenetic significance of this molecular character is unclear, our analyses do not support the close relationship between Cnidaria and Placozoa suggested by previous studies.
Assuntos
Cnidários/genética , DNA Ribossômico/genética , Animais , Estrutura Molecular , Filogenia , Análise de SequênciaAssuntos
DNA Satélite/genética , Genética Populacional , Macropodidae/genética , Alelos , Animais , Sequência de Bases , Primers do DNA , Variação Genética , Heterozigoto , Macropodidae/fisiologia , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Sequências Repetitivas de Ácido NucleicoRESUMO
Systematic relationships among the subclasses of Anthozoa, and especially among the orders Scleractinia, Actiniaria, and Corallimorpharia of subclass Zoantharia, were investigated by applying parsimony and distance methods of analysis to nucleotide sequence data obtained for the 5' end of the 28S rDNA. Exhaustive parsimony analysis indicates that the Ceriantipatharia are most representative of the ancestral Anthozoa. When applied to a wide range of scleractinians (nine taxa), actiniarians (seven taxa), and corallimorpharians (six taxa), both parsimony and distance analyses resolve three groups, one being the Scleractinia and the others containing both actiniarians and corallimorpharians. This indicates an unclear relationship between Actiniaria and Corallimorpharia and gives no support for Hand's hypothesis of scleractinian ancestry of actiniarians and corallimorphians; the monophyly of the Scleractinia, which is strongly supported by our analyses, is evidence to the contrary.
Assuntos
Cnidários/genética , DNA Ribossômico/genética , Filogenia , RNA Ribossômico 28S/genética , Animais , Sequência de Bases , Cnidários/classificação , Dados de Sequência Molecular , Homologia de Sequência do Ácido NucleicoRESUMO
We report the nucleotide sequence of the core histone gene cluster from the Cnidarian Acropora formosa. This is the first histone gene cluster to be sequenced from a diploblastic organism and the predicted amino acid sequences most resemble those of sea urchin equivalents. Each of the Cnidarian histone genes has two conserved regions 3' of the coding sequences and these closely resemble those of the metazoan alpha-class histone genes. In A. formosa the core histone genes are arranged as opposed (H3/H4 and H2A/H2B) pairs, a pattern common to the nondeuterostome metazoa, and tandem repetition is the predominant pattern of organization in the Cnidarian. With the recent identification of several classes of homeobox genes in Cnidarians these features clearly align the Cnidaria with triploblastic metazoans, supporting a monophyletic origin of the metazoa.