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Journal of Integrative Plant biology 2013年第11期55卷 1002-1015页

作者： Jiaojiao Wang Yi Zhang Jing Wu Lei Meng Haiyun Ren The key laboratory of cell proliferation and regulation biology of Ministry of Education College of Life Science Beijing Normal University

Formins are well-known regulators that participate in the organization of the actin cytoskeleton in organisms. The Arabidopsis thaliana L. genome encodes 21 formins, which can be divided into two distinct subfamilies. However, type II formins have to date been less well characterized. Here, we cloned a type II formin, AtFH16, and characterized its biochemical activities on actin and microtubule dynamics. The results show that the FH1 FH2 structure of AtFH16 cannot nucleate actin polymerization efficiently, but can bind and bundle microfilaments. AtFH16 FHIFH2 is also able to bind and bundle microtubules, and preferentially binds microtubules over microfilaments in vitro, in addition, AtFH16 FHIFH2 co-localizes with microtubules in onion epidermal cells, indicating a higher binding affinity of AtFH16 FHIFH2 for microtubules rather than microfilaments in vivo. In conclusion, AtFH16 is able to interact with both microfilaments and microtubules, suggesting that AtFH16 probably functions as a bifunctional protein, and may thus participate in plant cellular processes.

关键词： Actin filament Arabidopsis cytoskeleton formin microtubule.

Effect of PKCα on the gene expression related with proliferation and activity of transcription factor AP-1

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Chinese Science Bulletin 1997年第23期42卷 1986-1989页

作者： WANG Xiangyang and LIU Huitukey laboratory of cell proliferation and regulation biology, Department of biology, Beijing Normal University, Beijing 100875, China key laboratory of cell proliferation and regulation biology Department of Biology Beijing Normal University Beijing China

PROTEIN kinase C (PKC) is a serine/thronine kinase, which plays a major role in cell sig-nalling, growth control and tumorigenesis. PKC is encoded by a multigene family, which havebeen identified as consisting of at least 12 members in mammalian tissues. PKC isoforms differin their biochemical properties, tissue-specific expression and intracellular localization. Thedifferences imply that the individual PKC isoforms have a distinct physiological function. Inorder to study the role of specific PKC subtype in cell proliferation and transformation, weconstructed the human embryonic lung cell (2BS) model stably overexpressing PKC_α andfound that the overexpression of PKC_α promoted the growth rate and caused the transform-

关键词： protein kinase Cα 2BS cell transcription factor AP-1.

Construction of human embryonic lung cell model for overproduction of protein kinase Ca and its preliminary analysis

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Chinese Science Bulletin 1996年第6期41卷 506-511页

作者：王向阳吴承军柳惠图 State key laboratory of cell proliferation and regulation biology Department of Biology Beijing Normal University Beijing 100875 China

Protein kinase Cα（PKC）, a multifunctional Ca2+- and phospholipid-dependent kinase isthough to play an important role in cellular signal transduction, growth regulation andtumorigenesis. Molecular cloning technique demonstrated tha PKC was encoded by amultigene family. To date, at least 12 members of the PKC family have been *** is suggested that PKC isoforms differ in their cellular functions. In order to study thefurther role of specific PKC subtype in cell growth regulation, we constructed PKCα ex-

关键词： protein kinase Cα 2BS cell proliferation.

Actin Polymerization Mediated by AtFH5 Directs the Polarity Establishment and Vesicle Trafficking for Pollen Germination in Arabidopsis

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Molecular Plant 2018年第11期11卷 1389-1399页

作者： Chang Liu Yi Zhang Haiyun Ren key laboratory of cell proliferation and regulation biology of Ministry of Education College of Life ScienceBeijing Normal UniversityBeijing 100875China

The process of pollen germination is crucial for flowering plant reproduction,but the mechanisms through which pollen grains establish polarity and select germination sites are not well *** this study,we report that a formin family protein,AtFH5,is localized to the vesicles and rotates ahead of Lifeact-mEGFPlabeled actin filaments during pollen *** translocation of AtFH5 to the plasma membrane initiates the assembly of a collar-like actin structure at the prospective germination site prior to germination. Genetic and pharmacological evidence further revealed an interdependent relationship between the mobility of AtFH5-1 abeled vesicles and the polymerization of actin filaments:vesicle-localized AtFH5 promotes actin assembly,and the polymerization and elongation of actin filaments,in turn,is essential for the mobility of AtFH5-1 abeled vesicles in pollen *** together,our work revealed a molecular mechanism underlying the polarity establishment and vesicle mobility during pollen germination.

关键词： Chinese cabbage Pollen germination Polarity establishment Vesicle trafficking Actin Formin

The Villin/Gelsolin/Fragmin Superfamily Proteins in Plants

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Journal of Integrative Plant biology 2007年第8期49卷 1183-1191页

作者： Hui Su Ting Wang Huaijian Dong Haiyun Ren (key laboratory of cell proliferation and regulation biology of Ministry of Education and College of Life Sciences Beijing Normal University Beijing 100875 China key laboratory of cell proliferation and regulation biology of Ministry of Education and College of Life Sciences Beijing Normal University Beijing 100875 China

The villin/gelsolin/fragmin superfamily is a conserved Ca^2＋-dependent family of actin-regulating proteins that is widely present both in mammalian and non-mammalian organisms. They have traditionally been characterized by the same core of three or six tandem gelsolin subdomains. The study in vertebrates and lower eukaryotic cells has revealed that the villin/gelsolin/fragmin superfamily of proteins has versatile functions including severing, capping, nucleating or bundling actin filaments. In plants, encouraging progress has been made in this field of research in recent years. This review will summarize the identified plant homologs of villin/gelsolin/fragmin superfamily, thus providing a basis for reflection on their biochemical activities and functions in plants.

关键词： actin-binding protein actin cytoskeleton Villin/Gelsolin/Fragmin.

LISR28 Is Involved in Pollen Germination by Affecting Filamentous Actin Dynamics

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Molecular Plant 2013年第4期6卷 1163-1175页

作者： Li-Juan Cao Meng-Meng Zhao Chang Liu Huai-Jian Dong Wang-Cheng Li Hai-Yun Ren key laboratory of cell proliferation and regulation biology of Ministry of Education College of Life Science Beijing Normal University Beijing 100875China

Alternative splicing plays important roles in gene regulation and contributes to protein complexity. Previous studies suggest that alternative splicing exists in members of the villin/gelsolin/fragmin superfamily. In this study, a ser- ine/argine-rich （SR） protein cDNA with 28kDa protein （LISR28） was isolated from a lily （Lilium Iongiflorum） expression library. Protein domain analysis showed that LISR28 had similar structures to Arabidopsis SR45 （AtSR45）, and LISR28 could complement the phenotype of loss of AtSR45 function. Therefore, overexpression of LISR28 and AtSR45 mutant （atsr45-1） were used in the following experiments. Overexpression of LISR28 in Arabidopsis completely inhibited pollen germina- tion. In contrast, the pollen germination of atsr45-1 was earlier than that of wild-type. In addition, pollen of atsr45-1 contained less F-actin at the corresponding hydration stage during pollen germination compared to that of wild-type. Alternative splicing analysis showed that Arabidopsis villinl （AtVLN1） transcript encoding the full-length protein was increased, and that encoding the truncated protein was decreased in atst45-1. Moreover, the mRNA expression level of other actin-binding proteins （ABPs） abundant in Arabidopsis pollen was also changed in atsr45-1. In conclusion, we hypothesize that LISR28 alters F-actin dynamics probably through its alternative splicing activities to affect directly or indirectly the alternative splicing of AtVLN1 and the expression of different ABPs, which then affects the pollen germination.

关键词： SR protein alternative splicing F-actin.

MAPK signal pathways in the regulation of cell proliferation in mammalian cells

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cell Research 2002年第1期12卷 9-18页

作者： WEI ZHANG, Hui Tu LIU The key laboratory of cell proliferation and regulation biology of Ministry of Education, College of Life Sciences, Beijing Normal University, Beijing 100875, China 1The key laboratory of cell proliferation and regulation biology of Ministry of Education College of Life Sciences Beijing Normal University Beijing 100875 China 1The key laboratory of cell proliferation and regulation biology of Ministry of Education College of Life Sciences Beijing Normal University Beijing 100875 China

MAPK families play an important role in complex cellular programs like proliferation, differentiation, development, transformation, and apoptosis. At least three MAPI(families have been characterized: extracellular signal-regulated kinase (ERK), Jun kinase (JNK/SAPK) and p38 MAPK. The above effects are fulfilled by regulation of cell cycle engine and other cell proliferation related proteins. In this paper we discussed their functions and cooperation with other signal pathways in regulation of cell proliferation.

关键词： MAPK ER.K JNK/SPAK p38 cell proliferation cross-talk.

Effects of overexpressing p14^(ARF) on the apoptosis in human melanoma cells irradiated with γ-ray

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Chinese Science Bulletin 2003年第18期48卷 1971-1975页

作者： PENG Lixia, ZHANG Wei, LIU Huitu, HE Dacheng & GAO Ping key Laboratry of cell proliferation and regulation biology of Ministry of Education, College of Life Science, Beijing Normal University, Bei-jing 100875, China Correspondence should be addressed to Liu Huitu (e-mail: liuhuitu@ ***) key laboratory of cell proliferation and regulation biology of Ministry of Education College of Life Sciences Beijing Normal University

Tumor suppressor ARF can induce cell cycle arrest or apoptosis by activating p53. In order to explore the molecular mechanism of the induction of apoptosis by p14ARF, a human melanoma cell model overexpressing p14ARF was constructed. Present study indicated that in the cells overex-pressing p14ARF, p53 was accumulated in nucleus while it dispersed in cytosol in the control cells. Irradiated with g-ray, overexpressing p14ARF promoted the apoptosis of A375 cells, triggered Smac release from mitochondria to cytosol, and increased the expression of p53, Bax, Caspase-3, Caspase-9, p21cip1 and p27kip1. However, the protein level of Bcl-2 and phospho-ERK was down-regulated. These results suggested a possible mechanism of p14ARF in promotion of apoptosis.

关键词： p14ARF p53 g-ray apoptosis A375 cells.

Potential existence of two independent centrosome-targeting domains in PP4

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Chinese Science Bulletin 2007年第21期52卷 2930-2937页

作者： HUANG XiuQing NING LiFeng LONG ZhiTao SUN LingLing ZHANG Hui LIAN MuLan SANG JianLi key laboratory for cell proliferation and regulation biology of Ministry of Education College of Life Science Beijing NormalUniversity Beijing 100875 China

Protein phosphatase 4 (PP4) is an important member in the PPP family of protein Ser/Thr phosphatases. It has been proven to regulate a variety of cellular processes such as centrosome maturation, micro- tubule nucleation, splicesome assembly, and JNK pathway activation. Compared to the crystallized and structurally well defined phosphatase PP1 and PP2B, little is known about the structure of PP4. Besides the conserved motifs characteristic of the PPP family, no information is available on the other domains of PP4. PP4 is reported to localize to the centrosome in many species such as Drosophila, Caenor- habditis elegans and mammalian cells, which suggests a conserved role of PP4 in the regulation of centrosome function. Unlike several other centrosomal proteins, no sequence has been identified for PP4 that can target it to specific centrosomal localization. In this study, we used a combination of PCR mutagenesis and transient expression of GFP-tagged proteins in mammalian cells, and identified two PP4 centrosome-targeting domains of 68―136 and 134―220 aa. These two domains may be associated for appropriate localization to the centrosome. The findings are useful for further elucidating the func- tion of the domains and other structural characteristics of PP4.

关键词： PP4 细胞中心体局部化蛋白磷酸酶

Effects of PP4 suppression on the proliferation of MCF7 cells

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Chinese Science Bulletin 2006年第19期51卷 2335-2341页

作者： NING Lifeng LONG Zhitao HUANG Xiuqing SUN Lingling SANG Jianli The key laboratory for cell proliferation and regulation biology of Ministry of Education College of Life Science Beijing Normal University Beijing 100875

PP4, one of the few protein phosphata- ses associated with centrosome in cells of many species such as Drosophila, C. elegans and mam- mals, plays an essential role in the regulation of cen- trosome functions in Drosophila and C. elegans. In order to explore the role of PP4 in mammalian cells, full-length PP4 gene was obtained by RT-PCR from MCF7 cell total RNA and inserted into eukaryotic expression vector pEGFP-C1. The resultant con- struct pEGFP-C1-PP4 was transfected into MCF7 cells and immunostaining was carried out to confirm the centrosome localization of PP4. Then we re- versely subcloned a non-conserved domain of PP4 into pXJ41 to construct an anti-sense vector pXJ41- as-PP4. By transfecting pXJ41-as-PP4 into MCF7 cells and screening with G418, we obtained a stable cell line in which PP4 expression was stably sup- pressed. The cell line was analyzed on cell mor- phology, cytoskeleton structure, growth characteris- tics and the mitosis process. It was found that the proliferation rate decreased and serum-dependence increased in PP4-suppressed cells. Furthermore, flow cytometry and mitotic index analysis showed that G2/M transition was prolonged. PP4 suppression resulted in abnormal interphase microtubule, forma- tion of multipolar spindles and an increase in per- centage of multinuclear cells. These results sug- gested that PP4 is required for centrosome function in mammalian cells.

关键词：细胞周期 MCF7 PP4 蛋白质磷酸酶微管中心体有丝分裂反义RNA