Results: 5

1.
Figure 5

Figure 5. From: The protein kinase CK2 is involved in regulation of circadian rhythms in Arabidopsis.

Overexpression of CKB3 reduces phytochrome induction of Lhcb1*1. Northern analysis of Lhcb1*1 in 5-day-old etiolated plants after a 1-min exposure to red light. Closed squares, wild type; open circles, CKB3-ox (line ox41). Experiments were done twice with similar results.

Shoji Sugano, et al. Proc Natl Acad Sci U S A. 1999 October 26;96(22):12362-12366.
2.
Figure 4

Figure 4. From: The protein kinase CK2 is involved in regulation of circadian rhythms in Arabidopsis.

Periods of output genes are shortened in the CKB3-ox transgenic plants. Total RNA was isolated from wild-type and CKB3-ox (line ox18) plants grown as in Fig. 2, and expression of Lhcb1*1 (A), CCR2 (B), CAT2 (C), and CAT3 (D) RNAs was analyzed. Closed squares, wild type; open circles in red, CKB3-ox. The bar represents the subjective light conditions for the wild-type plants. Similar results were obtained in three experiments and with another CKB3-ox (ox41) line.

Shoji Sugano, et al. Proc Natl Acad Sci U S A. 1999 October 26;96(22):12362-12366.
3.
Figure 3

Figure 3. From: The protein kinase CK2 is involved in regulation of circadian rhythms in Arabidopsis.

CK2 can interact with and phosphorylate LHY in vitro. (A) Interaction of LHY with CKB3 in vitro. The input lane represents 10% of the 35S-labeled CKB3 used, and the other lanes show the amount of bound CKB3. The arrow indicates the position of the CKB3 protein. Molecular size markers are given (Left) in kDa. (B) Phosphorylation of LHY by CK2 in vitro. GST (lane 1) or GST-LHY (lanes 2 and 3) was incubated with 280 ng of CKA1 (lanes 1 and 3) or without CKA1 (lane 2) in the presence of [γ-32P]ATP. The arrow indicates the position of the GST-LHY protein after electrophoresis. (C) CK2 β-subunits enhance phosphorylation of LHY by CKA1. GST alone (lanes 1 and 2) or GST-LHY (lanes 3–5) was incubated with 14 ng of CKA1 and 35 ng of CKB1 (lanes 1 and 4) or CKB3 (lanes 2 and 5) or without β-subunits (lane 3). The arrow indicates the position of the GST-LHY protein.

Shoji Sugano, et al. Proc Natl Acad Sci U S A. 1999 October 26;96(22):12362-12366.
4.
Figure 1

Figure 1. From: The protein kinase CK2 is involved in regulation of circadian rhythms in Arabidopsis.

Overexpression of CKB3 increases CK2 activity. (A) Quantitative reverse transcription–PCR analysis of CKB3 and UBQ10 transcript levels in wild-type (WT) and independent homozygous transgenic lines (ox18 and ox41) after 14 days’ growth under L:D 16:8 photoperiods. The UBQ10 transcript levels were used as an internal control. (B) Western blot analysis of myc-CKB3 levels in transgenic lines grown as in A. The c-myc-tagged CKB3 protein (arrow) was detected with anti-c-myc antibody. Other bands are nonspecific. Molecular size markers are given (Left) in kDa. (C) CK2 activity in plant extracts prepared from wild-type and CKB3-ox lines. Plants were grown for 20 days under continuous white light. The data shown are means of two independent experiments for each line, with the range of the measurements indicated.

Shoji Sugano, et al. Proc Natl Acad Sci U S A. 1999 October 26;96(22):12362-12366.
5.
Figure 2

Figure 2. From: The protein kinase CK2 is involved in regulation of circadian rhythms in Arabidopsis.

Overexpression of CKB3 shortens periods of CCA1 and LHY circadian oscillations. (A) Northern blot analysis of circadian oscillation of CCA1 transcripts in wild-type and CKB3-ox (line ox18) plants. Plants were grown for 12 days in L:D 12:12 photoperiods, then transferred to continuous light after lights-on of day 13. A representative autoradiogram is shown (Upper). (Lower) Quantitation of one experiment. Values were normalized to the lowest value of the wild-type samples. Closed squares, wild type; open circles in red, CKB3-ox. The bar represents the subjective light conditions for the wild-type plants. Experiments were performed three times with similar results. The same results were also obtained in another CKB3-ox line (ox41). (B) Circadian oscillation of LHY RNA in wild-type and CKB3-ox plants. LHY transcripts in the same RNA samples were analyzed as in A. (C) CCA1 protein levels in wild-type and CKB3-ox (line ox18) plants. Proteins extracted from the same tissue used for RNA preparation were analyzed by Western blotting and detected with anti-CCA1 antibody. The arrow and line indicate CCA1 and nonspecific crossreacting proteins, respectively. Experiments were done twice with similar results.

Shoji Sugano, et al. Proc Natl Acad Sci U S A. 1999 October 26;96(22):12362-12366.

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