Fibroblast growth factor 2 (FGF-2) is a novel substrate for arginine methylation by PRMT5

Abstract Fibroblast growth factor 2 (FGF-2) is expressed in isoforms of different molecular masses from one mRNA species by alternative start of translation. The higher molecular mass isoforms (FGF-221 and 23) contain an arginine-rich N-terminus organized in RG-motifs followed by the 18 kDa FGF-2 (FGF-218) core which is common to all isoforms. Both isoforms localize differentially to the nucleus. Here, we analyzed the nuclear localization of FGF-221. Surprisingly, the lack of one RG-motif in FGF-221 resulted in the nucleolar distribution characteristic of FGF-218. We have previously shown that 23 kDa FGF-2 (FGF-223) interacts specifically with the survival of motoneuron (SMN) protein, an assembly protein for small nuclear ribonucleoprotein particles. For this assembly, Sm-proteins methylated by protein arginine methyltransferase 5 (PRMT5) are required. In our study, we aimed to analyze whether FGF-223 is also a substrate for symmetrical methylation by PRMT5. We could confirm that both proteins exist in a common complex. Moreover, PRMT5 methylates FGF-223 in vitro, whereas mutated inactive PRMT5 does not. FGF-223 is therefore a new substrate of PRMT5. With regard to function, inhibition of methyltransferase activity in HEK293T cells leads to cytoplasmic enrichment of FGF-2, indicating the importance of arginine methylation for shuttling of FGF-223 to the nucleus.

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Dynamic changes of podocytes caused by fibroblast growth factor 2 in culture

Cell and Tissue Research ◽

10.1007/s00441-021-03511-x ◽

2021 ◽

Author(s):

Eishin Yaoita ◽

Masaaki Nameta ◽

Yutaka Yoshida ◽

Hidehiko Fujinaka

Keyword(s):

Growth Factor ◽

Fibroblast Growth Factor ◽

Fibroblast Growth Factor 2 ◽

Quiescent State ◽

Fibroblast Growth ◽

Gene Expressions ◽

Dynamic Changes ◽

Ki 67

AbstractFibroblast growth factor 2 (FGF2) augments podocyte injury, which induces glomerulosclerosis, although the mechanisms remain obscure. In this study, we investigated the effects of FGF2 on cultured podocytes with interdigitating cell processes in rats. After 48 h incubation with FGF2 dynamic changes in the shape of primary processes and cell bodies of podocytes resulted in the loss of interdigitation, which was clearly shown by time-lapse photography. FGF2 reduced the gene expressions of constituents of the slit diaphragm, inflections of intercellular junctions positive for nephrin, and the width of the intercellular space. Immunostaining for the proliferation marker Ki-67 was rarely seen and weakly stained in the control without FGF2, whereas intensely stained cells were frequently found in the presence of FGF2. Binucleation and cell division were also observed, although no significant increase in cell number was shown. An in vitro scratch assay revealed that FGF2 enhanced migration of podocytes. These findings show that FGF2 makes podocytes to transition from the quiescent state into the cell cycle and change their morphology due to enhanced motility, and that the culture system in this study is useful for analyzing the pathological changes of podocytes in vivo.

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Human vascular injury triggers fibroblast growth factor 2 (FGF 2) release. Neointima formation in vitro is inhibited by novel catalytic DNA molecules targeting human EGR-1

Heart Lung and Circulation ◽

10.1046/j.1443-9506.2000.0622x.x ◽

2000 ◽

Vol 9 (3) ◽

pp. A95

Author(s):

H.C. Lowe ◽

M. Costandi ◽

C.N. Chesterman ◽

L.M. Khachigian

Keyword(s):

Growth Factor ◽

Fibroblast Growth Factor ◽

Vascular Injury ◽

Fibroblast Growth Factor 2 ◽

Neointima Formation ◽

Fibroblast Growth ◽

Dna Molecules ◽

Catalytic Dna

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Nuclear fibroblast growth factor-2 interacts specifically with splicing factor SF3a66

Biological Chemistry ◽

10.1515/bc.2004.156 ◽

2004 ◽

Vol 385 (12) ◽

pp. 1203-1208 ◽

Cited By ~ 13

Author(s):

Susanne Gringel ◽

Jeroen van Bergeijk ◽

Kirsten Haastert ◽

Claudia Grothe ◽

Peter Claus

Keyword(s):

Growth Factor ◽

Fibroblast Growth Factor ◽

Splicing Factor ◽

Fibroblast Growth Factor 2 ◽

Interacting Proteins ◽

Fibroblast Growth ◽

Extracellular Signaling ◽

C Terminus ◽

Assembly Factor ◽

Smn Protein

Abstract Fibroblast growth factor 2 (FGF-2) has a dual role as a classical extracellular signaling protein and as an intracellular factor. Isoforms of FGF-2, resulting from alternatively used start codons on one mRNA species, locate differentially to nuclear compartments. In this study we aimed to analyze functions of intracellular FGF-2 by identification of interacting proteins. We identified the 66-kDa subunit of splicing factor 3a (SF3a66) as a binding partner in a yeast two-hybrid screen and confirmed this interaction by pull-down assays. The splicing factor interacted with the 18-kDa (FGF-218) and with the 23-kDa (FGF-223) isoforms, indicating an interaction with a domain common to both isoforms. Moreover, FGF-2 interacted with the C-terminus of SF3a66, a sequence that has not previously been assigned a functional role. In a functional neurite outgrowth assay, SF3a66 enhanced neurite lengths similar to FGF-218. We have previously identified the spliceosomal assembly factor survival of motoneuron (SMN) protein as a protein interacting specifically with the FGF-223 isoform [Claus et al., J. Biol. Chem. 278 (2003), 479–485]. The identification of two FGF-2 interacting proteins from the same biochemical pathway suggests a novel intranuclear role of FGF-2.

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A New 34-Kilodalton Isoform of Human Fibroblast Growth Factor 2 Is Cap Dependently Synthesized by Using a Non-AUG Start Codon and Behaves as a Survival Factor

Molecular and Cellular Biology ◽

10.1128/mcb.19.1.505 ◽

1999 ◽

Vol 19 (1) ◽

pp. 505-514 ◽

Cited By ~ 146

Author(s):

Emmanuelle Arnaud ◽

Christian Touriol ◽

Christel Boutonnet ◽

Marie-Claire Gensac ◽

Stéphan Vagner ◽

...

Keyword(s):

Growth Factor ◽

Fibroblast Growth Factor ◽

Human Fibroblast ◽

The Other ◽

In Vitro Translation ◽

Start Codon ◽

Fibroblast Growth Factor 2 ◽

Initiation Codon ◽

Fibroblast Growth

ABSTRACT Four isoforms of human fibroblast growth factor 2 (FGF-2) result from alternative initiations of translation at three CUG start codons and one AUG start codon. Here we characterize a new 34-kDa FGF-2 isoform whose expression is initiated at a fifth initiation codon. This 34-kDa FGF-2 was identified in HeLa cells by using an N-terminal directed antibody. Its initiation codon was identified by site-directed mutagenesis as being a CUG codon located at 86 nucleotides (nt) from the FGF-2 mRNA 5′ end. Both in vitro translation and COS-7 cell transfection using bicistronic RNAs demonstrated that the 34-kDa FGF-2 was exclusively expressed in a cap-dependent manner. This contrasted with the expression of the other FGF-2 isoforms of 18, 22, 22.5, and 24 kDa, which is controlled by an internal ribosome entry site (IRES). Strikingly, expression of the other FGF-2 isoforms became partly cap dependent in vitro in the presence of the 5,823-nt-long 3′ untranslated region of FGF-2 mRNA. Thus, the FGF-2 mRNA can be translated both by cap-dependent and IRES-driven mechanisms, the balance between these two mechanisms modulating the ratio of the different FGF-2 isoforms. The function of the new FGF-2 was also investigated. We found that the 34-kDa FGF-2, in contrast to the other isoforms, permitted NIH 3T3 cell survival in low-serum conditions. A new arginine-rich nuclear localization sequence (NLS) in the N-terminal region of the 34-kDa FGF-2 was characterized and found to be similar to the NLS of human immunodeficiency virus type 1 Rev protein. These data suggest that the function of the 34-kDa FGF-2 is mediated by nuclear targets.

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Localized delivery of fibroblast growth factor–2 and brain-derived neurotrophic factor reduces spontaneous seizures in an epilepsy model

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.0810710106 ◽

2009 ◽

Vol 106 (17) ◽

pp. 7191-7196 ◽

Cited By ~ 96

Author(s):

Beatrice Paradiso ◽

Peggy Marconi ◽

Silvia Zucchini ◽

Elena Berto ◽

Anna Binaschi ◽

...

Keyword(s):

Growth Factor ◽

Fibroblast Growth Factor ◽

Neurotrophic Factor ◽

Viral Vectors ◽

Neuronal Damage ◽

Brain Derived Neurotrophic Factor ◽

Fibroblast Growth Factor 2 ◽

Fibroblast Growth ◽

Damage Limitation

A loss of neurons is observed in the hippocampus of many patients with epilepsies of temporal lobe origin. It has been hypothesized that damage limitation or repair, for example using neurotrophic factors (NTFs), may prevent the transformation of a normal tissue into epileptic (epileptogenesis). Here, we used viral vectors to locally supplement two NTFs, fibroblast growth factor–2 (FGF-2) and brain-derived neurotrophic factor (BDNF), when epileptogenic damage was already in place. These vectors were first characterized in vitro, where they increased proliferation of neural progenitors and favored their differentiation into neurons, and they were then tested in a model of status epilepticus-induced neurodegeneration and epileptogenesis. When injected in a lesioned hippocampus, FGF-2/BDNF expressing vectors increased neuronogenesis, embanked neuronal damage, and reduced epileptogenesis. It is concluded that reduction of damage reduces epileptogenesis and that supplementing specific NTFs in lesion areas represents a new approach to the therapy of neuronal damage and of its consequences.

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