Multifocal two-photon laser scanning microscopy combined with photo-activatable GFP for in vivo monitoring of intracellular protein dynamics in real time

Martini, Joerg; Schmied, Katja; Palmisano, Ralf; Tœnsing, Katja; Anselmetti, Dario; Merkle, Thomas

doi:10.1016/j.jsb.2006.12.012

Cited by 23 publications

(19 citation statements)

References 25 publications

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“…To test this possibility, HMGB2 and HMGB4 fused to photoactivatable GFP (paGFP) were expressed in tobacco protoplasts and examined by multifocal two-photon laser scanning microscopy (Martini et al, 2007). The dynamics of the diffusion of HMGB2 and HMGB4 fused to paGFP was recorded before (0 s) and at different times during and after activation of the paGFP in the nucleus (Fig.…”

Section: Hmgb2 and Hmgb4 Shuttle Between The Nucleus And The Cytoplasmmentioning

confidence: 99%

“…For expression of GFP-HMG fusions in stably transformed Arabidopsis plants, the GFP-HMG coding sequences were inserted into plasmid pGII0179-35S (Launholt et al, 2006). For analysis of protein dynamics, the HMGB coding sequences were inserted with paGFP in plasmid p5#paGFP (Martini et al, 2007). For expression of the wild-type and mutant forms of HMGB2 in Escherichia coli, the coding sequences were inserted into plasmid pQE9cm (Grasser et al, 1996), providing an N-terminal 63 His tag.…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…The analysis of real-time protein mobility in transiently transformed tobacco BY-2 protoplasts expressing HMGB proteins fused to paGFP using multifocal two-photon laser scanning microscopy was performed as described previously (Martini et al, 2007).…”

Section: Monitoring Of Intracellular Protein Dynamics Using Pagfpmentioning

confidence: 99%

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Nucleocytoplasmic Distribution of the Arabidopsis Chromatin-Associated HMGB2/3 and HMGB4 Proteins

et al. 2010

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High mobility group (HMG) proteins of the HMGB family are chromatin-associated proteins that as architectural factors are involved in the regulation of transcription and other DNA-dependent processes. HMGB proteins are generally considered nuclear proteins, although mammalian HMGB1 can also be detected in the cytoplasm and outside of cells. Plant HMGB proteins studied so far were found exclusively in the cell nucleus. Using immunofluorescence and fluorescence microscopy of HMGB proteins fused to the green fluorescent protein, we have examined the subcellular localization of the Arabidopsis (Arabidopsis thaliana) HMGB2/3 and HMGB4 proteins, revealing that, in addition to a prominent nuclear localization, they can be detected also in the cytoplasm. The nucleocytoplasmic distribution appears to depend on the cell type. By time-lapse fluorescence microscopy, it was observed that the HMGB2 and HMGB4 proteins tagged with photoactivatable green fluorescent protein can shuttle between the nucleus and the cytoplasm, while HMGB1 remains nuclear. The balance between the basic amino-terminal and the acidic carboxyl-terminal domains flanking the central HMG box DNA-binding domain critically influences the nucleocytoplasmic distribution of the HMGB proteins. Moreover, protein kinase CK2-mediated phosphorylation of the acidic tail modulates the intranuclear distribution of HMGB2. Collectively, our results show that, in contrast to other Arabidopsis HMGB proteins such as HMGB1 and HMGB5, the HMGB2/3 and HMGB4 proteins occur preferentially in the cell nucleus, but to various extents also in the cytoplasm.Within the cell nucleus, the genomic DNA is organized with histones and other proteins into a nucleoprotein complex termed chromatin. This packaging of the DNA has crucial consequences for DNA-dependent processes, including the transcription of genes. The chromatin structure is highly dynamic and is modulated by a variety of chromatin-associated proteins. Among these proteins are the high mobility group (HMG) proteins that represent a heterogeneous class of proteins that, after the histones, are the second most abundant family of chromosomal proteins

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Section: Hmgb2 and Hmgb4 Shuttle Between The Nucleus And The Cytoplasmmentioning

confidence: 99%

Section: Plasmid Constructionmentioning

confidence: 99%

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Nucleocytoplasmic Distribution of the Arabidopsis Chromatin-Associated HMGB2/3 and HMGB4 Proteins

et al. 2010

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“…3G-I). By contrast, colocalisation analysis of the endoplasmic reticulum/Golgi marker Pra1-DsRed(Martini et al, 2007) and HMGB1-GFP demonstrates different localisation of the two fusion proteins(Fig. 3J-L).…”

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confidence: 89%

“…3J-L). For comparison, the localisation of a cytosolic marker GFP-NLS(mut)-CHS-NES(Haasen et al, 1999) and a nuclear marker NLS-CHS-DsRed(Martini et al, 2007) are shown(Fig. 3M and …”

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confidence: 99%

Chromosomal high mobility group (HMG) proteins of the HMGB-type occurring in the moss Physcomitrella patens

Kiilerich

Stemmer²,

Merkle

et al. 2008

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Computed tomography screening for lung cancer: Review of screening principles and update on current status

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2007

Cancer

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Screening for lung cancer with low-dose computed tomography (CT) is controversial. In favor of screening, lung cancer is the leading cause of cancer death in the United States, and those at greatest risk are identified readily on the basis of age and smoking history. In addition, it is well established that CT is far more sensitive than chest radiography in detecting lung cancer when it is small and asymptomatic. Furthermore, very high rates of survival were reported recently for screen-detected lung cancers in a large, multinational, single-arm observational study. However, a reduction in lung cancer mortality has not been demonstrated to date, and a recent longitudinal study with a simulated control group suggested little or no mortality reduction. In addition, there are important harms from CT screening, including false-positive test results and overdiagnosis. Furthermore, healthcare resources are finite. Therefore, even if the benefits do outweigh the harms, the cost-effectiveness of CT screening for lung cancer still will need to be considered in the context of competing healthcare alternatives. The objectives of this article were 3-fold: 1) to review the basic principles of screening and study designs related to cancer screening, 2) to summarize the results of the observational and analytical studies of CT screening that have been reported to date, and 3) to describe the design of the 2 ongoing, randomized controlled trials of CT screening and what may be learned from these studies in the near future. Cancer

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Multifocal two-photon laser scanning microscopy combined with photo-activatable GFP for in vivo monitoring of intracellular protein dynamics in real time

Cited by 23 publications

References 25 publications

Nucleocytoplasmic Distribution of the Arabidopsis Chromatin-Associated HMGB2/3 and HMGB4 Proteins

Nucleocytoplasmic Distribution of the Arabidopsis Chromatin-Associated HMGB2/3 and HMGB4 Proteins

Chromosomal high mobility group (HMG) proteins of the HMGB-type occurring in the moss Physcomitrella patens

Computed tomography screening for lung cancer: Review of screening principles and update on current status

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