Differential Regulation of the AP-1 Family Members by UV Irradiation In Vitro and In Vivo

The LASS (longevity assurance homologue) family members are highly conserved from yeasts to mammals. Five mouse and human LASS family members, namely LASS1, LASS2, LASS4, LASS5 and LASS6, have been identified and characterized. In the present study we cloned two transcriptional variants of hitherto-uncharacterized mouse LASS3 cDNA, which encode a 384-amino-acid protein (LASS3) and a 419-amino-acid protein (LASS3-long). In vivo, [3H]dihydrosphingosine labelling and electrospray-ionization MS revealed that overproduction of either LASS3 isoform results in increases in several ceramide species, with some preference toward those having middle- to long-chain-fatty acyl-CoAs. A similar substrate preference was observed in an in vitro (dihydro)ceramide synthase assay. These results indicate that LASS3 possesses (dihydro)ceramide synthesis activity with relatively broad substrate specificity. We also found that, except for a weak display in skin, LASS3 mRNA expression is limited almost solely to testis, implying that LASS3 plays an important role in this gland.

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Short-Form Bomanins Mediate Humoral Immunity in Drosophila

Journal of Innate Immunity ◽

10.1159/000489831 ◽

2018 ◽

Vol 10 (4) ◽

pp. 306-314 ◽

Cited By ~ 14

Author(s):

Scott A. Lindsay ◽

Samuel J.H. Lin ◽

Steven A. Wasserman

Keyword(s):

Humoral Immunity ◽

Short Form ◽

Family Members ◽

Gene Encoding ◽

Vitro Assay ◽

Gram Positive Bacteria ◽

Toll Receptor ◽

Secreted Peptides

The Bomanins (Boms) are a family of a dozen secreted peptides that mediate the innate immune response governed by the Drosophila Toll receptor. We recently showed that deleting a cluster of 10 Bom genes blocks Toll-mediated defenses against a range of fungi and gram-positive bacteria. Here, we characterize the activity of individual Bom family members. We provide evidence that the Boms overlap in function and that a single Bom gene encoding a mature peptide of just 16 amino acids can act largely or entirely independent of other family members to provide phenotypic rescue in vivo. We further demonstrate that the Boms function in Drosophila humoral immunity, mediating the killing of the fungal pathogen Candida glabrata in an in vitro assay of cell-free hemolymph. In addition, we find that the level of antifungal activity both in vivo and in vitro is linked to the level of Bom gene expression. Although Toll dictates expression of the antimicrobial peptides (AMPs) drosomycin and metchnikowin, we find no evidence that Boms act by modifying the expression of the mature forms of these antifungal AMPs.

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Characterization of calcium- and integrin-binding protein 1 (CIB1) knockout platelets: Potential compensation by CIB family members

Thrombosis and Haemostasis ◽

10.1160/th08-06-0351 ◽

2008 ◽

Vol 100 (05) ◽

pp. 847-856 ◽

Cited By ~ 19

Author(s):

Brenda R. Temple ◽

Holly R. Gentry ◽

Jan C. DeNofrio ◽

Weiping Yuan ◽

Leslie V. Parise

Keyword(s):

Thrombus Formation ◽

Mrna Level ◽

Family Members ◽

Cytoplasmic Tail ◽

Binding Pocket ◽

Vitro Binding ◽

Cytoplasmic Tails ◽

Hydrophobic Binding

SummaryPlatelet aggregation requires activation of the αIIbβ3 integrin,an event regulated by the integrin cytoplasmic tails. CIB1 binds to the cytoplasmic tail of the integrin αIIb subunit. Previous overexpression and knockdown studies in murine megakaryocytes demonstrated that CIB1 inhibits integrin αIIbβ3 activation.Here we analyzed Cib1-/- mice to determine the function of CIB1 in platelets in vitro and in vivo. We found that although these mice had no overt platelet phenotype, mRNA level of CIB1 homolog CIB3 was increased in Cib1-/- megakaryocytes. In vitro binding experiments showed that recombinant CIB1, -2 and -3 bound specifically to an αIIb cytoplasmic tail peptide. Subsequent protein modeling experiments indicated that CIBs 1–3 each have a highly conserved hydrophobic binding pocket. Therefore, the potential exists for compensation for the loss of CIB1 by these CIB family members, thereby preventing pathologic thrombus formation in Cib1-/- mice.

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Differential regulation of detoxification enzymes in hepatic and mammary tissue by hops (Humulus lupulus ) in vitro and in vivo

Molecular Nutrition & Food Research ◽

10.1002/mnfr.201200534 ◽

2013 ◽

Vol 57 (6) ◽

pp. 1055-1066 ◽

Cited By ~ 25

Author(s):

Birgit M. Dietz ◽

Ghenet K. Hagos ◽

Jillian N. Eskra ◽

Gihani T. Wijewickrama ◽

Jeffrey R. Anderson ◽

...

Keyword(s):

Differential Regulation ◽

Humulus Lupulus ◽

Detoxification Enzymes ◽

Mammary Tissue

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Differential regulation of HSP27 oligomerization in tumor cells grown in vitro and in vivo

Oncogene ◽

10.1038/sj.onc.1203850 ◽

2000 ◽

Vol 19 (42) ◽

pp. 4855-4863 ◽

Cited By ~ 99

Author(s):

Jean-Marie Bruey ◽

Catherine Paul ◽

Annie Fromentin ◽

Sophie Hilpert ◽

André-Patrick Arrigo ◽

...

Keyword(s):

Tumor Cells ◽

Differential Regulation

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Bcl-2 family members in childhood acute lymphoblastic leukemia: relationships with features at presentation, in vitro and in vivo drug response and long-term clinical outcome

Leukemia ◽

10.1038/sj.leu.2401529 ◽

1999 ◽

Vol 13 (10) ◽

pp. 1574-1580 ◽

Cited By ~ 46

Author(s):

GS Salomons ◽

LA Smets ◽

M Verwijs-Janssen ◽

AAM Hart ◽

EG Haarman ◽

...

Keyword(s):

Acute Lymphoblastic Leukemia ◽

Clinical Outcome ◽

Drug Response ◽

Lymphoblastic Leukemia ◽

Family Members ◽

Childhood Acute Lymphoblastic Leukemia

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The Biological and Clinical Relevance of Inhibitor of Growth (ING) Genes in Non-Small Cell Lung Cancer

Cancers ◽

10.3390/cancers11081118 ◽

2019 ◽

Vol 11 (8) ◽

pp. 1118 ◽

Cited By ~ 2

Author(s):

Elisabeth Smolle ◽

Nicole Fink-Neuboeck ◽

Joerg Lindenmann ◽

Freyja Smolle-Juettner ◽

Martin Pichler

Keyword(s):

Lung Cancer ◽

Family Members ◽

In Vivo Studies ◽

Action Mode ◽

Inhibition Of Growth ◽

Molecular Mechanism Of Action ◽

Anchor Points ◽

Lung Cancer Detection

Carcinogenic mutations allow cells to escape governing mechanisms that commonly inhibit uncontrolled cell proliferation and maintain tightly regulated homeostasis between cell death and survival. Members of the inhibition of growth (ING) family act as tumor suppressors, governing cell cycle, apoptosis and cellular senescence. The molecular mechanism of action of ING genes, as well as their anchor points in pathways commonly linked to malignant transformation of cells, have been studied with respect to a variety of cancer specimens. This review of the current literature focuses specifically on the action mode of ING family members in lung cancer. We have summarized data from in vitro and in vivo studies, highlighting the effects of varying levels of ING expression in cancer cells. Based on the increasing insight into the function of these proteins, the use of ING family members as clinically useful biomarkers for lung cancer detection and prognosis will probably become routine in everyday clinical practice.

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The role of BH3-only protein Bim extends beyond inhibiting Bcl-2–like prosurvival proteins

The Journal of Cell Biology ◽

10.1083/jcb.200905153 ◽

2009 ◽

Vol 186 (3) ◽

pp. 355-362 ◽

Cited By ~ 131

Author(s):

Delphine Mérino ◽

Maybelline Giam ◽

Peter D. Hughes ◽

Owen M. Siggs ◽

Klaus Heger ◽

...

Keyword(s):

Family Members ◽

Binding Specificity ◽

In Vitro Studies ◽

Genetic Approach ◽

Activation Model ◽

Bh3 Domain ◽

Indirect Activation

Proteins of the Bcl-2 family are critical regulators of apoptosis, but how its BH3-only members activate the essential effectors Bax and Bak remains controversial. The indirect activation model suggests that they simply must neutralize all of the prosurvival Bcl-2 family members, whereas the direct activation model proposes that Bim and Bid must activate Bax and Bak directly. As numerous in vitro studies have not resolved this issue, we have investigated Bim's activity in vivo by a genetic approach. Because the BH3 domain determines binding specificity for Bcl-2 relatives, we generated mice having the Bim BH3 domain replaced by that of Bad, Noxa, or Puma. The mutants bound the expected subsets of prosurvival relatives but lost interaction with Bax. Analysis of the mice showed that Bim's proapoptotic activity is not solely caused by its ability to engage its prosurvival relatives or solely to its binding to Bax. Thus, initiation of apoptosis in vivo appears to require features of both models.

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NF-κB/Rel family members are physically associated phosphoproteins

Biochemical Journal ◽

10.1042/bj3030499 ◽

1994 ◽

Vol 303 (2) ◽

pp. 499-506 ◽

Cited By ~ 38

Author(s):

C C H Li ◽

M Korner ◽

D K Ferris ◽

E Chen ◽

R M Dai ◽

...

Keyword(s):

Phorbol Ester ◽

Family Members ◽

Jurkat Cells ◽

Nf Kappa B ◽

I Kappa B Alpha ◽

Jurkat T Cell ◽

Phosphopeptide Mapping ◽

First Time

We performed radioimmunoprecipitation followed by serial immunoblots to show that, in the unstimulated Jurkat T cell line, the NF-kappa B/Rel family proteins, p80-c-Rel, p105-NF-kappa B, p65-NF-kappa B, p50-NF-kappa B and p36-I kappa B alpha, can be detected as complexes using antisera against c-Rel, p105-NF-kappa B or p65-NF-kappa B. p36-I kappa B alpha and p105, both known inhibitors of NF-kappa B function, can physically associate with NF-kappa B/Rel family members, but not with each other. In vivo and in vitro phosphorylation experiments demonstrated that NF-kappa B/Rel family members, including p105, c-Rel, p50, p65 (for the first time for p50 and p65) and p36-I kappa B alpha are also phosphoproteins. Phosphoserine and phosphothreonine residues were identified in these proteins isolated from unstimulated Jurkat cells. Both unphosphorylated and hyperphosphorylated forms of p36-I kappa B alpha were found in the complexes, suggesting that hyperphosphorylated I kappa B alpha is still capable of associating with the NF-kappa B/Rel family members. After stimulation with phorbol 12-myristate 13-acetate and phytohaemagglutinin for 10 min, p105-NF-kappa B and p50-NF-kappa B, but not p36-I kappa B, were highly phosphorylated. Phosphopeptide mapping of p105 showed that phorbol ester/phytohaemagglutinin stimulation may change p105 phosphorylation qualitatively.

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