Annual Report 2006

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and interleukin (IL)-2 synthesis in the T-cell receptor (TCR) signaling pathway. The anti- WASP-EVH1 scFvs were constructed with a VH leader signal sequence at the N-terminus and with a light chain constant (CL) region behind the VL region (Fig. 1B), and injected into fertilized mouse eggs. In T cells from transgenic mice overexpressing the anti-WASP -EVH1 scFvs, binding of anti-WASP-EVH1 scFvs to native WASP was confirmed by immunoprecipitation. Furthermore, IL-2 production induced by TCR stimulation was strongly inhibited in the scFv transgenic mice T cells (Fig. 2A). Similar T-cell defects were observed in WASP-EVH1 domain overexpressing transgenic mice (Fig. 2A). So, the WASP-EVH1 scFv intrabodies inhibit only the function of EVH1 domain that regulates IL-2 synthesis signaling, but do not affect the rest of the domain functions of WASP (Fig. 2B). The novel procedure presented here can be applicable to functional analysis of other cytosolic proteins. Fig. 2 IL-2 production induced by TCR stimulation was inhibited by anti-WASP-EVH1 intrabodies (A) Splenic T cells from anti-WASP scFv 21SHL and 21SHL-CL Tg mice, WASP-EVH1 Tg and wild-type mice were cultured in medium alone or in the presence of anti-CD3εAb. Each cell culture supernatant was collected at 24 h. IL-2 in the supernatant was quantified by ELISA. (B) Schematic representation of mode of intrabody action against WASP-EVH1 domain in T cells. WASP contains multiple domains that enable it to interact with different proteins. Shown are the EVH1 domain, GTPase-binding domain (GBD), proline rich region (PRR), verprolin homology (VH) and cofilin homology (CH) domain. Anti-WASP scFvs can specifically bind to the EVH1 domain of WASP in the cytosol of T cells, blocking the collect interaction between WASP and signaling molecules involved in IL-2 synthesis.
Mulberry latex defense mulberry trees from herbivorous insects Kotaro KONNO Insect Genetics and Evolution Department Latex is widely found among plant species; 12,000-35,000 species have been reported to exude it and many kinds of chemicals and proteins have been reported from plant latex. The biological functions of plant latex and its ingredients, however, remained obscure. Recently, we found that several latex-producing plants with no reported toxicities are strongly toxic to insects due to the ingredients of latex. For example, papaya ( Moraceae) and fig ( Moraceae) leaves are strongly toxic to insects because of the cysteine proteases in their latex. Our recent findings indicated a need to investigate latex ingredients and their biological functions. Mulberry trees ( Moraceae) grow in Asia and their leaves are used for rearinganeconomicallyveryimportantinsect, the silkworm, , for thousands of years. Moraceae plants are characterized by the presence of latex, and mulberry trees also exude latex (Fig. 1, lower left photo) when their leaves are damaged by caterpillars. In wild condition, mulberry leaves are not often damaged by herbivorous insects, although the leaves are soft and contains a lot of nutrients Fig. 1 Defense activity of mulberry latex and three sugar-mimic alkaloids, the active compounds contained in the latex in very high concentrations Upper left photo, left part: the cabbage moth larvae fed intact (excised) mulberry leaves; Upper left photo, right part: the cabbage moth larvae fed mulberry leaves from which latex was washed of by cutting into narrow leaf strips and washing with water; Lower left photo: latex exuded from mulberry leaves(arrows), Right: three sugar-mimic alkaloids contained in mulberry latex in high concentrations, which are glycosidase inhibitors and were reported to have antidiabetic activities.
Page 1 and 2: 2006
Page 3 and 4: Teruo Ishige, President The Nationa
Page 5 and 6: List of Publication Original paper
Page 7 and 8: Topics of Research in This Year Pos
Page 9 and 10: studies. To construct a fully satur
Page 11 and 12: Fig. 2 Geographical distribution of
Page 13: (ABR0015, ABR0257, ABR0417, ABR0495
Page 17 and 18: Fig. 2 Growth inhibitory effects of
Page 19 and 20: Fig. 3 Change of the levels of luci
Page 21 and 22: Next, we examined the effect of tra
Page 23 and 24: G enome Research Department Introdu
Page 25 and 26: The Rice Annotation Project Databas
Page 27 and 28: industrial and agricultural concern
Page 29 and 30: One of the main problems involving
Page 31 and 32: Fig. 3 Mapping of QTL for rachis in
Page 33 and 34: two genes, XOO2263 and XOO4134 cont
Page 35 and 36: G enebank Genebank Project The NIAS
Page 37 and 38: using dormant winter buds. Also, we
Page 39 and 40: Insect and Animal Sciences Division
Page 41 and 42: Expression patterns of two novel ge
Page 43 and 44: Fig. 5 PGC and EBC fused cells PGCs
Page 45 and 46: cells. The epidermal cells maintain
Page 47 and 48: Animal and cell culture models for
Page 49 and 50: terminus, respectively. A signal se
Page 51 and 52: Fig. 2 Trehalose content of body pa
Page 53 and 54: Fig. 6 Birth weight of 11 adult SNT
Page 55 and 56: The role of glucose as a metabolic
Page 57 and 58: analysis using microsatellite marke
Page 59 and 60: Fig. 2 Recorded Electromyograms (le
Page 61 and 62: Table 1 Menu of Commercial silkworm
Page 63 and 64: Fig. 3 Expression of GFP gene in th
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also surveyed in the genetic stocks
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esearch were located in or near gen
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elatives results in severe reductio
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and circadian clocks may control th
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B iochemistry Department Structural
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Proteomic analysis Based on the pro
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photoautotrophic medium (Fig. 1B).
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Fig. 4 Blast resistance of OsWRKY45
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phosphatase activity by SIPK depend
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the endosperm in co-operation with
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isolated genes in transgenic Anal
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Fig. 1 Gene structures and mutation
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cytochimera among the mulberries (
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15 G. Hariraj, Kinoshita Haruo, T.
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49 Kameda Tsunenori, Teramoto Hidet
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85 Matsuyama Shuichi, Ohkura Satosh
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120 Ozawa Kenjiro, Kawahigashi Hiro
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155 Taguchi-Shiobara Fumio, Yamamot
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192 Ueno Osamu, Agarie Sakae, (2005
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8 Murakami Ritsuko, Koyama Akio, Sh
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20 Todoroki Junichi, Kaneko Hiroyuk
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Executive Members and Research Staf
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Developmental Biology Department Di
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Research Leader Hayasaka Shoji Rese
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Mutation Genetics Laboratory Head N
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thousands of yen TOTAL BUDGET OPERA
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Annual Report 2006 (Apr. 2005Mar. 2
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