Consecutive Steps of Phosphorylation Affect Conformation and DNA Binding of the Chironomus High Mobility Group A Protein

Schwanbeck, Ralf; Gymnopoulos, Marco; Petry, Inga; Piekiełko, Agnieszka; Szewczuk, Zbigniew; Heyduk, Tomasz; Zechel, Kasper; Wiśniewski, Jacek R.

doi:10.1074/jbc.m011053200

Cited by 25 publications

(26 citation statements)

References 53 publications

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“…Indeed, the ability to tweak both protein stability and conformation of the intrinsically unstructured HMGA proteins by covalent modifications such as phosphorylation or acetylation underlies their participation in diverse biological processes from transcription to recombination (18,62). The C-terminal HMGA-like part of CarD would be similarly malleable to such conformational and so functional alterations.…”

Section: Discussionmentioning

confidence: 99%

“…Protein conformational changes and decreases in DNA binding affinity are also brought about by phosphorylation by a number of kinases including CKII, Cdc2 kinase, mitogen-activated protein kinase, and protein kinase C (14 -18). This provokes fluctuations in intracellular protein stability and in DNA binding, thereby fine-tuning their regulatory functions in vivo (16,18).…”

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confidence: 99%

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Domain Architecture of a High Mobility Group A-type Bacterial Transcriptional Factor

Padmanabhan

Elías‐Arnanz

Carpio

et al. 2001

Journal of Biological Chemistry

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Myxococcus xanthus transcriptional factor CarD participates in carotenogenesis and fruiting body formation. It is the only reported prokaryotic protein having adjacent "AT-hook" DNA-binding and acidic regions characteristic of eukaryotic high mobility group A (HMGA) proteins. The latter are small, unstructured, nonhistone nuclear proteins that function as architectural factors to remodel DNA and chromatin structure and modulate various DNA binding activities. We find CarD to be predominantly dimeric with two stable domains: (a) an N-terminal domain of defined secondary and tertiary structure which is absent in eukaryotic HMGA proteins; (b) a C-terminal domain formed by the acidic and AT-hook segments and lacking defined structure. CarD, like HMGA proteins, binds specifically to the minor-groove of AT-rich DNA present in two appropriately spaced tracts. As in HMGA proteins, casein kinase II can phosphorylate the CarD acidic region, and this dramatically decreases the DNA binding affinity of CarD. The acidic region, in addition to modulating DNA binding, confers structural stability to CarD. We discuss how the structural and functional plasticity arising from domain organization in CarD could be linked to its role as a general transcriptional factor in M. xanthus.

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Section: Discussionmentioning

confidence: 99%

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Domain Architecture of a High Mobility Group A-type Bacterial Transcriptional Factor

Padmanabhan

Elías‐Arnanz

Carpio

et al. 2001

Journal of Biological Chemistry

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“…Further research revealed that HMGA1 proteins were phosphorylated by several kinases such as cyclindependent kinase 1 (previously known as cdc2 kinase) [12,13], protein kinase C (PKC) [14], and protein kinase CK2 [15]. These phosphorylations attenuated the DNA binding affinities of mammalian HMGA1a [14,16,17] and HMGA1b [17,18].…”

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A quantitative study on the in vitro and in vivo acetylation of high mobility group A1 proteins

Zhang

Yan

et al. 2007

J. Am. Soc. Mass Spectrom.

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High mobility group (HMG) A1 proteins are subject to a number of post-translational modifications, which may regulate their function in gene transcription and other cellular processes. We examined, by using mass spectrometry, the acetylation of HMGA1a and HMGA1b proteins induced by histone acetyltransferases p300 and PCAF in vitro and in PC-3 human prostate cancer cells in vivo. It turned out that five lysine residues in HMGA1a, i.e., Lys-14, Lys-64, Lys-66, Lys-70, and Lys-73, could be acetylated by both p300 and PCAF. We further quantified the level of acetylation by analyzing, with LC-MS/MS, the proteolytic peptides of the in vitro or in vivo acetylated HMGA1 proteins where the unmodified lysine residues were chemically derivatized with a perdeuterated acetyl group. Quantification results revealed that p300 and PCAF exhibited different site preferences for the acetylation; the preference of p300 acetylation followed the order of Lys-64ϳLys-70 Ͼ Lys-66 Ͼ Lys-14ϳLys73, whereas the selectivity of PCAF acetylation followed the sequence of Lys-70ϳLys-73 Ͼ Lys-64ϳLys-66 Ͼ Lys-14. HMGA1b was acetylated in a very similar fashion as HMGA1a. We also demonstrated that C-terminal phosphorylation of HMGA1 proteins did not affect the in vitro acetylation of the two proteins by either p300 or PCAF. Moreover, we examined the acetylation of lysine residues in HMGA1a and HMGA1b isolated from PC-3 human prostate cancer cells. Our results showed that all the above five lysine residues were also acetylated in vivo, with Lys-64, Lys-66 and Lys-70 in HMGA1a exhibiting higher levels of acetylation than Lys-14 and Lys-73. (J Am Soc Mass Spectrom 2007, 18, 1569 -1578

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“…The AT hooks have random structure when free but adopt a defined conformation on binding specifically to the narrow minor groove of AT-rich sequences 4 to 8 bp in length and present in at least two appropriately spaced tracts (20,25,43). Kinases such as casein kinase II (CKII) and Cdc2 phosphorylate HMGA proteins and modulate DNA binding as well as protein stability (14,32,36,41,47,49,50). This occurs in a cell cycle-and differentiationdependent manner and fine-tunes HMGA activity in vivo.…”

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The Stigmatella aurantiaca Homolog of Myxococcus xanthus High-Mobility-Group A-Type Transcription Factor CarD: Insights into the Functional Modules of CarD and Their Distribution in Bacteria

et al. 2003

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Transcriptional factor CarD is the only reported prokaryotic analog of eukaryotic high-mobility-group A (HMGA) proteins, in that it has contiguous acidic and AT hook DNA-binding segments and multifunctional roles in Myxococcus xanthus carotenogenesis and fruiting body formation. HMGA proteins are small, randomly structured, nonhistone, nuclear architectural factors that remodel DNA and chromatin structure. Here we report on a second AT hook protein, CarD Sa , that is very similar to CarD and that occurs in the bacterium Stigmatella aurantiaca. CarD Sa has a C-terminal HMGA-like domain with three AT hooks and a highly acidic adjacent region with one predicted casein kinase II (CKII) phosphorylation site, compared to the four AT hooks and five CKII sites in CarD. Both proteins have a nearly identical 180-residue N-terminal segment that is absent in HMGA proteins. In vitro, CarD Sa exhibits the specific minor-groove binding to appropriately spaced AT-rich DNA that is characteristic of CarD or HMGA proteins, and it is also phosphorylated by CKII. In vivo, CarD Sa or a variant without the single CKII phosphorylation site can replace CarD in M. xanthus carotenogenesis and fruiting body formation. These two cellular processes absolutely require that the highly conserved N-terminal domain be present. Thus, three AT hooks are sufficient, the N-terminal domain is essential, and phosphorylation in the acidic region by a CKII-type kinase can be dispensed with for CarD function in M. xanthus carotenogenesis and fruiting body development. Whereas a number of hypothetical proteins homologous to the N-terminal region occur in a diverse array of bacterial species, eukaryotic HMGA-type domains appear to be confined primarily to myxobacteria.

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Consecutive Steps of Phosphorylation Affect Conformation and DNA Binding of the Chironomus High Mobility Group A Protein

Cited by 25 publications

References 53 publications

Domain Architecture of a High Mobility Group A-type Bacterial Transcriptional Factor

Domain Architecture of a High Mobility Group A-type Bacterial Transcriptional Factor

A quantitative study on the in vitro and in vivo acetylation of high mobility group A1 proteins

The Stigmatella aurantiaca Homolog of Myxococcus xanthus High-Mobility-Group A-Type Transcription Factor CarD: Insights into the Functional Modules of CarD and Their Distribution in Bacteria

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