Faculty Profiles - SUZUKI Nobuhiro

LOW-TEMPERATURE-DEPENDENT EXPRESSION OF A RICE GENE ENCODING A PROTEIN WITH A LEUCINE-ZIPPER MOTIF

K AGUAN, K SUGAWARA, N SUZUKI, T KUSANO

MOLECULAR & GENERAL GENETICS 240 ( 1 ) 1 - 8 1993.7

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER VERLAG

We have isolated from rice suspension cells three non-sequence-related cDNAs the expression of which is markedly induced by low, non-freezing temperature. Here we further characterize one of the cDNA clones, lip19. Expression of lip19 is positively regulated by low temperature, but not affected by high (40-degrees-C) temperature. Sequencing and primer extension analyses showed that lip19 has a long (552 bp) 5' non-coding sequence followed by a single open reading frame specifying a protein of 148 amino acids. The deduced amino acid sequence of the protein, Lip19, shows at its amino-terminus a conserved basic region followed by a ''leucine-zipper'' domain. The reported sequence most similar to Lip19 is maize OCSBF-1, which is a bZip-type DNA binding protein. The possibility is suggested that Lip19 is a transcriptional factor that is positively controlled by low temperature.

DOI： 10.1007/BF00276876

Molecular analysis of rice dwarf phytoreovirus segment S1: Interviral homology of the putative RNA-dependent RNA polymerase between plant- and animal-infecting reoviruses

Nobuhiro Suzuki, Masako Tanimura, Yuzuko Watanabe, Tomonobu Kusano, Yoshichika Kitagawa, Narushi Suda, Hiroshi Kudo, Ichiro Uyeda, Eishiro Shikata

Virology 190 ( 1 ) 240 - 247 1992

Language：English Publishing type：Research paper (scientific journal)

We have determined the complete nucleotide sequence of the largest segment S1 of rice dwarf phytoreovirus (RDV), a member of the family Reoviridae. S1 is 4423 nucleotides long with a segment-specific inverted repeat located adjacent to the conserved termini (5′GGCAAA- - -UGAU3′). A major open reading frame (bases 36 to 4367) on the S1 plus strand, which is preceded by a minicistron (bases 6 to 29), encodes the polypeptide (P1) consisting of 1444 amino acids with a Mr of 164, 142. The sense-strand transcript derived from the full-length S1 cDNA, the minicistron of which was abolished, directed the synthesis of a polypeptide of 170 kDa in addition to smaller polypeptides in wheat germ extracts, and the 170-kDa product comigrated with the minor core protein in SDS-polyacrylamide gel. Thus, P1 is assumed to be localized in the viral core particle. The consensus sequence element conserved in RNA-dependent RNA polymerase is observed in the P1 amino acid sequence predicted from the nucleotide sequence. Based on the dendrogram established from the sequence alignment around the polymerase module region, and sequence identity within the alignment, P1 of plant-infecting RDV was evolutionarily compared with VP1, λ3, and VP1 of three other animal-infecting members of the family, rota-, reo-, and bluetongue viruses. Consequently, RDV S1 was shown to be more closely related to the rotavirus gene segment 1, in terms of molecular evolution, than the animal-infecting members are to one another. © 1992.

DOI： 10.1016/0042-6822(92)91210-L

Rice dwarf phytoreovirus segment S12 transcript is tricistronic in Vitro

Nobuhiro Suzuki, Mikiko Sugawara, Tomonobu Kusano

Virology 191 ( 2 ) 992 - 995 1992

Language：English Publishing type：Research paper (scientific journal)

Sequence analysis revealed that rice dwarf phytoreovirus segment S12 is 1066 nucleotides long with a small out-of-phase, overlapping open reading frame (ORF) as well as a major ORF. The large ORF (positions 42 to 980) encodes 312 amino acids, while the small one (bases 313 to 591) encodes 92 amino acids with an additional in-frame AUG codon (positions 337-339) 24 nucleotides downstream from the first one. Transcripts from a full-length cDNA directed the in vitro synthesis of three polypeptides of 33 (considered to be translated from the long ORF), 8, and 7 kDa. Alteration of each of the two ATG codons on the small ORF demonstrated their involvement in the generation of the 8- and 7-kDa polypeptides. Although it is still unknown whether these proteins are expressed in vivo, the small ORF is shown to be conserved in S9s of two other members of the genus Phytoreovirus, rice gall dwarf virus and wound tumor virus, suggesting its common, important function. © 1992.

DOI： 10.1016/0042-6822(92)90279-X

ISOLATION OF GENES FOR LOW-TEMPERATURE-INDUCED PROTEINS IN RICE BY A SIMPLE SUBTRACTIVE METHOD

K AGUAN, K SUGAWARA, N SUZUKI, T KUSANO

PLANT AND CELL PHYSIOLOGY 32 ( 8 ) 1285 - 1289 1991.12

Language：English Publishing type：Research paper (scientific journal) Publisher：JAPANESE SOC PLANT PHYSIOLOGISTS

We have isolated cDNAs that correspond to three distinct low-temperature-induced mRNAs from rice cells using an effective, simple subtraction method. The corresponding genes were designated lip (gene encoding low-temperature-induced protein) -5, -9 and -19. The expression of these genes was slightly stimulated (lip5 and lip19) or unaffected (lip9) by abscisic acid, unlike the expression of the rab16A-D genes which are readily induced by either abscisic acid or high osmotic stress. Time-course analysis revealed that while the lip5 gene was induced 3 h after temperature down-shift, and the lip9 and lip19 genes were induced 6 h after such a shift.

Web of Science

MOLECULAR ANALYSIS OF RICE DWARF PHYTOREOVIRUS SEGMENT-S11 CORRESPONDING TO WOUND TUMOR PHYTOREOVIRUS SEGMENT-S12 Reviewed

N SUZUKI, M HARADA, T KUSANO

JOURNAL OF GENERAL VIROLOGY 72 2233 - 2237 1991.9

Language：English Publishing type：Research paper (scientific journal) Publisher：SOC GENERAL MICROBIOLOGY

The complete nucleotide sequence of rice dwarf phytoreovirus (RDV) genome segment S11 was determined. S11 is 1067 nucleotides long. There is an inverted repeat of 10 bp adjacent to the conserved 5'-terminal hexanucleotide (5' GGUAAA 3') and 3'-terminal tetranucleotide (5' UAGU 3') sequences. A single large open reading frame found in the plus strand of S11 begins with the first AUG codon (bases 6 to 8) and extends for 567 bases. Evolutionary relatedness between RDV S11 and wound tumour phytoreovirus S12 based on amino acid sequence similarity (25.8%) was found. In addition to the first AUG triplet, RDV S11 possesses a second in-phase AUG triplet (positions 30 to 32) nearby, which conforms to the Kozak consensus sequence. Two forms of the protein were identified by using an in vitro transcription and translation system in which a tailored full-length cDNA was the initial template. The abolition of the first AUG codon by site-directed mutagenesis resulted in disappearance of the larger translation product. These results strongly suggest that the two products are translated from the first and second AUG codons. Whether the two proteins are expressed in vivo is at present unclear.

Web of Science

MOLECULAR-CLONING AND EXPRESSION OF THIOBACILLUS-FERROOXIDANS CHROMOSOMAL RIBULOSE BISPHOSPHATE CARBOXYLASE GENES IN ESCHERICHIA-COLI

T KUSANO, K SUGAWARA, C INOUE, N SUZUKI

CURRENT MICROBIOLOGY 22 ( 1 ) 35 - 41 1991.1

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER VERLAG

The genes coding for d-ribulose-1,5-bisphosphate carboxylase (RuBPCase) from an iron-oxidizing bacterium, Thiobacillus ferrooxidans, were cloned into an Escherichia coli plasmid, pUC18. The recombinant plasmid, termed pTR11, contained a 4.0-kb PstI fragment including the entire coding regions for both large and small subunits of RuBPCase. Escherichia coli carrying pTR11 did not show any CO2-fixing activity. However, a derivative plasmid with an appropriate deletion, which was placed under the control of a tac promoter, conferred ribulose bisphosphate-dependent CO2-fixing activity on the host cell. Analysis of gel-filtration chromatography of the RuBPCase synthesized in E. coli revealed that it had a hexadecameric form like the native enzyme of T. ferrooxidans. © 1991 Springer-Verlag New York Inc.

DOI： 10.1007/BF02106210

Web of Science

Outer capsid protein heterogeneity of rice dwarf phytoreovirus

N. Suzuki, M. Sugawara

Journal of General Virology 72 ( 9 ) 2239 - 2242 1991

Language：English Publishing type：Research paper (scientific journal)

The 46K outer capsid protein encoded by RNA segment S8 and the 42K polypeptide, previously thought to be the segment S9-encoded structural protein, were isolated from a rice dwarf phytoreovirus purified preparation, and then analysed by peptide mapping and electroblot-ELlSA. Staphylococcus aureus V8 protease peptide mapping patterns of the 42K and 46K proteins were similar. Two monoclonal antibodies (MAbs), obtained after immunization with virus particles dissociated by 0.1% SDS, were each specific for both the 42K and 46K proteins. Furthermore, the MAbs bound common peptide fragments which were generated by digestion of the 42K and 46K proteins with V8 protease or proteinase K. These results strongly suggest that the 42K protein is not a gene product of S9 but a product overlapping with the 46K outer capsid protein. Whether the two proteins are functionally distinct remains to be determined.

DOI： 10.1099/0022-1317-72-9-2239

Molecular analysis of rice dwarf phytoreovirus segment S11 corresponding to wound tumour phytoreovirus segment S12

N. Suzuki, M. Harada, T. Kusano

Journal of General Virology 72 ( 9 ) 2233 - 2237 1991

Language：English Publishing type：Research paper (scientific journal)

The complete nucleotide sequence of rice dwarf phytoreovirus (RDV) genome segment S11 was determined. S11 is 1067 nucleotides long. There is an inverted repeat of 10 bp adjacent to the conserved 5'terminal hexanucleotide (5' GGUAAA 3') and 3'terminal tetranucleotide (5' UAGU 3') sequences. A single large open reading frame found in the plus strand of S11 begins with the first AUG codon (bases 6 to 8) and extends for 567 bases. Evolutionary relatedness between RDV S11 and wound tumour phytoreovirus S12 based on amino acid sequence similarity (25-8%) was found. In addition to the first AUG triplet, RDV S11 possesses a second in-phase AUG triplet (positions 30 to 32) nearby, which conforms to the Kozak consensus sequence. Two forms of the protein were identified by using an in vitro transcription and translation system in which a tailored full-length cDNA was the initial template. The abolition of the first AUG codon by site-directed mutagenesis resulted in disappearance of the larger translation product. These results strongly suggest that the two products are translated from the first and second AUG codons. Whether the two proteins are expressed in vivo is at present unclear.

DOI： 10.1099/0022-1317-72-9-2233

AN IMPROVED METHOD FOR THE CONSTRUCTION OF HIGH-EFFICIENCY CDNA LIBRARY IN PLASMID OR LAMBDA VECTOR Reviewed

K AGUAN, T KUSANO, N SUZUKI, Y KITAGAWA

NUCLEIC ACIDS RESEARCH 18 ( 4 ) 1071 - 1071 1990.2

ISOLATION AND SEROLOGICAL COMPARISON OF VIRUS-CODED PROTEINS OF 3 POTYVIRUSES INFECTING CUCURBITACEOUS PLANTS Reviewed

N SUZUKI, Y SHIRAKO, Y EHARA

INTERVIROLOGY 31 ( 1 ) 43 - 49 1990.1

Language：English Publishing type：Research paper (scientific journal) Publisher：KARGER

Cylindrical inclusion proteins (CIPs), amorphous inclusion proteins (AIPs) and virus particles were partially purified from Cucurbita maxima leaves infected with zucchini yellow mosaic virus (ZYMV), watermelon mosaic virus 2 (WMV2), and the watermelon mosaic virus 1 strain of papaya ringspot virus (PRSV-W). Antisera to these individual antigens, except AIPs of ZYMV and WMV2, were prepared and used to determine with electroblot-ELISA the serological differentiation indices (SDIs) between homologous and heterologous antigens. The average SDIs between ZYMV and WMV2 CIPs, between ZYMV and PRSV-W CIPs, and between WMV2 and PRSV-W CIPs were 1.5, 3.5, and 2.5, respectively. The SDIs between ZYMV and PRSV-W AIPs and between WMV2 and PRSV-W AIPs were 5 and 3, respectively. The average SDIs between ZYMV and WMV2 capsid proteins (CPs), between ZYMV and PRSV-W CPs, and between WMV2 and PRSV-W CPs were 3, 7, and 8, respectively. These data suggest that the serological relationship between ZYMV and WMV2 is much closer than that between PRSV-W and ZYMV or between PRSV-W and WMV2 and that antigenic determinants of CIP and AIP were conserved more than those of CP among the three potyviruses.

DOI： 10.1159/000150133

Sequence analysis of rice dwarf phytoreovirus genome segments S4, S5, and S6: Comparison with the equivalent wound tumor virus segments Reviewed

Nobuhiro Suzuki, Yuzuko Watanabe, Tomonobu Kusano, Yoshichika Kitagawa

Virology 179 ( 1 ) 446 - 454 1990

Language：English Publishing type：Research paper (scientific journal)

The complete nucleotide sequences of genome segments S4, S5, and S6 of rice dwarf phytoreovirus (RDV) were determined. S4 and S5 consist of 2468 and 2570 base pairs, respectively, S5 thus being larger in size than S4, contrary to the situation suggested by their relative migration in a polyacrylamide gel. S6 is 1699 nucleotides long. The individual segments have segment-specific inverted repeats adjacent to the conserved terminal sequences (5′GGUAAA-UGAU3′ for S4, 5′GGCAAA-UGAU3′ for S5 and S6). S4, S5, and S6 each have single long open reading frames encoding 727, 801, and 509 amino acids, respectively. A low level of amino acid sequence homology was observed between RDV S4 and wound tumor virus (WTV) S4 (22.4%), and between RDV S6 and WTV S6 (20.2%). On the other hand, RDV S5 and WTV S5 show 52.0% amino acid sequence similarity, indicating that S5 is much more conserved than any other segments of RDV and WTV reported so far. Further comparative analyses indicate that the RDV segment shows a greater frequency of usage of codons XYG and XYC, and much less frequent usage of codon XYA than the equivalent WTV segment, this codon preference bias being more conspicuous than expected from the base contents. © 1990.

DOI： 10.1016/0042-6822(90)90313-G

Sequence analysis of the rice dwarf phytoreovirus segment s3 transcript encoding for the major structural core protein of 114 kDa Reviewed

Nobuhiro Suzuki, Yuzuko Watanabe, Tomonobu Kusano, Yoshichika Kitagawa

Virology 179 ( 1 ) 455 - 459 1990

Language：English Publishing type：Research paper (scientific journal)

The primary structure of rice dwarf phytoreovirus (RDV) genome segment S3 was determined. RDV S3 consists of 3195 nucleotides. A 14-bp segment-specific inverted repeat is located immediately adjacent to the conserved terminal sequence (5′GGCAAA---UGAU3′). A single long open reading frame encoding for 1019 amino acids with an Mr of 114,289 is also identified. In order to investigate the localization of the predicted polypeptide, we determined the amino acid sequence of the 26-kDa peptide fragment obtained from the structural core protein digested by Staphylococcus aureus V8 protease. The sequence of the fragment was found in the translational product presumed from the nucleotide sequence of RDV S3, indicating that RDV S3 encodes the major structural core protein of 114 kDa. © 1990.

DOI： 10.1016/0042-6822(90)90314-H