The N-terminal region of RECQL4 lacking the helicase domain is both essential and sufficient for the viability of vertebrate cells. Role of the N-terminal region of RECQL4 in cells.
Rothmund-Thomson syndrome (RTS) is a rare genetic disorder characterized by premature aging, developmental abnormalities, and a predisposition to cancer. RTS is caused by mutations in the RECQL4 gene, which encodes one of the five human RecQ helicases. To identify the cellular functions of RECQL4, we generated a chicken DT40 cell line in which RECQL4 expression could be turned off by doxycycline (Dox). Upon exposure to Dox, cells stopped growing and underwent apoptosis. The cells could be rescued by expression of the N-terminal region of RECQL4 (amino acids 1-496), which lacks the helicase domain and has sequence similarity to yeast Sld2, which plays an essential function in the initiation of DNA replication in Saccharomyces cerevisiae. Smaller fragments of the N-terminal region of RECQL4 did not rescue the cells from lethality. RECQL4 gene knockout cells complemented with RECQL4 (1-496) showed relatively high sensitivity to DNA damaging agents that induce double strand breaks and cross-links, suggesting that the C-terminal region including the helicase domain of RECQL4 is involved in the repair of certain types of DNA lesions.
Molecular Cell Biology Laboratory, Graduate School of Pharmaceutical Sciences, Tohoku University, Aoba 6-3, Aramaki, Aoba-ku, Sendai 980-8578, Japan.
Takuya
T
Abe
Akari
A
Yoshimura
Yoshifumi
Y
Hosono
Shusuke
S
Tada
Masayuki
M
Seki
Takemi
T
Enomoto
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D016923:Cell Death; D002460:Cell Line; D002470:Cell Survival; D002645:Chickens; D004265:DNA Helicases / Q000378:metabolism; D004260:DNA Repair; D004318:Doxycycline / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D055786:Gene Knockout Techniques; D006801:Humans; D017434:Protein Structure, Tertiary; D053484:RecQ Helicases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011038:Rothmund-Thomson Syndrome / Q000235:genetics* / Q000378:metabolism
D000900:Anti-Bacterial Agents; C506731:RECQL4 protein, human; D004265:DNA Helicases; D053484:RecQ Helicases; D004318:Doxycycline