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K Fujinaga

Publications and source records attributed to K Fujinaga.

At least 181 records · Page 10Linked to original sources
PubMed1982

Altered properties of tumors induced by adenovirus type 12 DNA fragment transformed cells after growth in immunocompetent rats.

The GY1-3-1 cell line, which was derived from a rat cell line transformed by the Ad 12 HindIII-G fragment, can induce tumors in newborn rats but not in mature rats. However, when 10 mature rats were transplanted with a large number of GY1-3-1 tumor fragments, two developed tumors with a long latency period. These two tumors (GY1-3M and GY1-3Y) were quite different in properties from the originally inoculated GY1-3-1 tumor. Tumor transplantation studies revealed that both GY1-3M and GY1-3Y tumors could grow in mature rats and the number of takes with transplanted tumor was 100% with GY1-3M tumor and 40% with GY1-3Y tumor. Histopathologically original GY1-3-1 tumor showed a morphology of undifferentiated sarcoma, while the histologic picture of GY1-3M and GY1-3Y tumors was similar to that encountered in human fibrosarcoma and malignant fibrous histiocytoma, respectively. Both GY1-3M and GY1-3Y tumors, when recultured in vitro, gave rise to cell lines with a fibroblastic appearance although the original GY1-3-1 cell line exhibited an epithelioid morphology. In chromosome analysis, the GY1-3-1 cell line was pseudodiploid, while the cell lines from GY1-3M and GY1-3Y tumors were hyperdiploid or near-triploid. The metacentric marker chromosome, MI, was present in mitotic cells of the GY1-3-1 line and of the GY1-3M tumor lines, but absent from the GY1-3Y tumor lines. By Southern blot hybridization, multiple bands of cellular DNAs from parental GY1-3-1 cells hybridized with labelled Ad 12 HindIII-G, while only a single band hybridized DNAs from both GY1-3M and GY1-3Y tumors.

Adenoviruses, Human↗
PubMed1982

A simple and practical method for typing and strain differentiation of herpes simplex virus using infected cell DNAs.

A simple and practical method for typing and strain differentiation of herpes simplex virus (HSV) isolates, based upon analysis of the restriction endonuclease cleavage patterns of viral DNAs, was established by using unlabeled infected cell DNAs. The preparation of infected cell DNA is technically easier than that of purified viral DNA or of radiolabeled viral DNA. The method provides a powerful and practical tool for epidemiological and clinical studies of HSV infection, which can be performed in most diagnostic laboratories. In order to select suitable restriction endonucleases for the study of HSV isolates, the cleavage patterns of viral DNAs (strains MacIntyre, HF, UW-268, and SAV) with 12 enzymes were analyzed. Several enzymes, Bam HI, Kpn I, Pst I, Sal I, Sst I, and Xho I, were found to be useful for both typing and strain differentiation. With this method, HSV isolates from different individuals and from the same individual were analyzed by digestion of their infected cell DNAs with Bam HI. Six isolates from epidemiologically unrelated individuals were readily typed and differentiated from each other. Three isolates from the same individual showed very similar patterns. However, there was a small degree of difference between the first two isolates and the third isolate.

DNA Restriction Enzymes↗
PubMed1982

Organization and expression of oncogenic human adenovirus transforming genes.

The transforming genes of the oncogenic human adenoviruses (Ad) reside within the left 7-8% of the viral genome. The organization and the expression of the entire Ad12 E1 region (the leftmost 11.2%) including the transforming genes, 3,860 nucleotide pairs, have been analyzed. Nucleotide sequencing and mRNA mapping studies on the entire E1 region made it possible to assign sites of mRNA initiations, splicings and terminations, and the regions coding for polypeptides at the nucleotide level. Amino acid sequences of the highly-oncogenic Ad12 E1 polypeptides thus predicted were compared with those of non-oncogenic Ad5, and the existence of a high degree of homologies in most of the Ad E1 polypeptides was shown between highly-oncogenic Ad12 and non-oncogenic Ad5. Among the viral-specific mRNAs identified, not only species corresponding to those synthesized in infected KB cells during early stages of productive infection, but also unique chimeric ones containing sequences of viral-viral or cellular-viral junctions were detected in cells transformed by Ad12 and Ad7 transforming genes. Some cDNA clones of the chimeric mRNA species as well as genomic DNA clones of the junction regions from Ad7 transformed cells were isolated and analyzed.

Adenoviruses, Human↗
PubMed1982

[Adenovirus].

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Adenoviruses, Human↗
PubMed1981

Nucleotide sequence of the transforming early region E1b of adenovirus type 12 DNA: structure and gene organization, and comparison with those of adenovirus type 5 DNA.

The nucleotide sequence of the entire transforming early region of E1b of the highly oncogenic adenovirus type 12 (Ad12) DNA has been determined. The total sequence (3860 base pairs) encompasses the entire transforming early region E1 of Ad12 DNA. From the sequence for the E1b region of Ad12, and the transcription map of the E1b region (1, 2, 3, and this paper) the structure and gene organization of the early region E1b of Ad12 DNA were analyzed and compared with those of the E1b region in the non-oncogenic Ad5 DNA (4, 5). Most of the sequences in the E1b region of Ad12 was highly homologous to that of Ad5. It is predicted that the Ad12 region E1b codes for polypeptides of 53.9, 19.1, and 8.9 kd. This situation is identical with that of the Ad5 region E1b which codes for polypeptides of 54.9, 20.6, and 8.3 kd. The function of these predicted polypeptides encoded by the E1b regions in cell transformation is discussed.

Adenoviridae↗
PubMed1981

Grouping of adenoviruses and identification of restriction endonuclease cleavage patterns of adenovirus DNAs using infected cell DNA: simple and practical methods.

Simple and practical methods for grouping of adenoviruses and for identification of restriction endonuclease cleavage patterns of viral DNA were established by using infected cell DNA. DNA homology groupings of adenoviruses could be examined by spot hybridization, and restriction endonuclease cleavage patterns of viral DNAs could be obtained by Southern blot hybridization, by using infected cell DNA. The method was very sensitive and allowed the identification of the cleavage pattern of viral DNA of the inoculum by means of cell DNA extracted from infected cells with undetectable cytopathic effect (CPE). In ethidium bromide-stained gels without Southern blot hybridization, the restriction endonuclease cleavage pattern of viral DNA could be detected precisely in spite of background staining due to cellular DNA. The preparation of infected cell DNA used in these procedures was technically much easier than that of viral DNA. These methods require only a small number of infected cells and allow many isolates to be investigated with ease.

Adenoviruses, Human↗
PubMed1980

Chromosomal alterations of rat cell lines transformed by human adenovirus type-12 virion, whole DNA and left-end DNA fragments.

A normal rat cell line, 3Y1-B clone 1-6 (3Y1) and its adenovirus (Ad) type-12-transformed derivatives, W4 (transformed by Ad12 Virion), WY3 (transformed by Ad12 whole DNA), CY1-1 (transformed by the Ad12 EcoRI-C fragment, left 16.5%), GY1-1 (transformed by the Ad12 HindIII-G fragment, left 6.8%) and HY1 (transformed by the Ad12 Acd-H fragment, left 4.7%) were studied cytogenetically. 3Y1 and some of the transformed cell lines (W4, WY3 and GY1-1) were diploid or pseudodiploid, while others (CY1-1 and HY1) were hypotetraploid. A metacentric marker M1 was detected in GY1-1 cells and another marker M2 in W4 and CY1-1 cells at a high frequency. By the Giemsa banding technique, the metacentric markers M1 and M2 from these fully transformed cell lines were identified as isochromosomes derived from 1q (M1) and 3q (M2), respectively. On the other hand, the markers were detected only at a low frequency in incompletely transformed HY1 cells. However, hypersomy in chromosome No. 1 was observed at a high frequency in this cell line. It can be concluded that hypersomy of chromosomes No. 1 or 3 found in transformants and metacentric markers found in complete transformants are characteristic features in rat cells transformed by Ad12.

Adenoviruses, Human↗
PubMed1980

Structure and gene organization in the transformed Hind III-G fragment of Ad12.

The nucleotide sequence of the transforming Hind III-G fragment of Ad12 DNA which encompasses the left 6.8% of the genome has been determined. The fragment was 2320 nucleotides long, and contained a GC cluster at positions 126-155 and a region extremely rich in AT at positions 1098-1142 (number from the leftmost end). Possible coding regions for the two transforming gene products were assigned. The predicted coding region for T antigen g is positions 502-1069 and positions 1144-1373, which are joined by splicing (266 amino acid residues, 30 kd), and that for T antigen f is positions 1845-2126 (94 amino acid residues, 10 kd). The sequence of the Hind III-G fragment was compared with that of the transforming DNA fragment of Ad5 which encompasses the left 8.0% of the genome (2809 nucleotides). There are several discrete regions with significant sequence homology. The comparison suggests that the regions in the left two thirds of the Ad5 and Ad12 transforming DNA fragments (map units 0-4.7% in Ad5 and 0-4.4% in Ad12) bear some resemblance in their gene organizations, and code for proteins containing structurally homologous regions.

Adenoviruses, Human↗
PubMed1980

Unique species of mRNA from adenovirus type 7 early region 1 in cells transformed by adenovirus type 7 DNA fragment.

Adenovirus type 7 (Ad7) early region 1 mRNA species transcribed in rat cell lines transformed by the HindIII-I . J fragment (the left 7.8% of the viral genome) and in human KB cells infected with Ad7 were mapped on the viral genome, using S1 nuclease gel and diazobenzyloxymethyl paper hybridization techniques. At the early stage of productive infection, two mRNA's (950 and 840 nucleotides long) with the common 5' and 3' ends but different internal splicings were mapped from region 1A (map units 1.4 to 4.3), and one mRNA (2,310 nucleotides long, with the internal splicing between map units 9.9 to 10.1) was mapped from region 1B (map units 4.6 to 11.4). At the late stage, these early spliced mRNA's were also found and at least three additional Ad7 mRNA's were identified: 700-nucleotide-long mRNA in region 1A; and 1,100- and nucleotide-long mRNA's in region 1B. In transformed rat cell lines, two early region 1A mRNA's (950 and 840 nucleotides long) were also transcribed. Surprisingly, in addition, several unique Ad7 mRNA's, not found in productivity infected cells, were identified in all of the transformed cell lines. Their molecular sizes and coding sequences varied in individual cell lines. However, these mRNA's had the 5' end-proximal portion in region 1B and the 3' end-proximal portion in region 1A, these portions being transcribed by extending from region 1B to 1A on viral DNA fragments joined in a tandem array in transformed cells.

Adenoviruses, Human↗
PubMed1980

Mapping of adenovirus 12 mRNA's transcribed from the transforming region.

Adenovirus 12 mRNA's transcribed from the transforming region were analyzed and mapped on viral DNA by the nuclease S1 gel and diazobenzyloxymethyl paper blot techniques in cells transformed of adenovirus 12 DNA and in cells early and late after lytic infection with adenovirus 12. Two initiation sites of mRNA transcription and two kinds of splicing were found in each of early regions 1A and 1B. For early region 1A mRNA's, four species were found in lytically infected cells. Three of them were commonly found in cells transformed by either HindIII-G or EcoRI-C. Cells transformed by HindIII-G contained two additional 1A transcripts, which could be the 3' portions of chimeric mRNA's of cellular-viral or viral-viral sequences. Transcription in the 1B region diverged among the above cell lines. Early after lytic infection, no appreciable amount of 1B mRNA was detected, whereas two species of mRNA's, one corresponding to protein IX mRNA and another having splicing, were found at the late stage. In cells transformed by EcoRI-C, a distinct mRNA species with splicing was observed. Cells transformed by HindIII-G contained a transcript from the leftmost part of the 1B sequence at the 5' portion of chimeric mRNA species, suggesting the presence of tandem integration of viral DNA in the cells. Other mRNA species in the 1A and 1B regions were also detected in both transformed cell lines. The results are discussed in relation to the nucleotide sequence of the HindIII-G fragment of adenovirus 12 DNA.

Adenoviruses, Human↗