A novel mutation in the juxtamembrane intracellular sequence of the granulocyte colony-stimulating factor (G-CSF) receptor gene in a patient with severe congenital neutropenia augments GCSF proliferation activity but not through the MAP kinase cascade.
We analyzed the structure of the granulocyte colony-stimulating factor (G-CSF) receptor gene in a 6-year-old female patient with severe congenital neutropenia (SCN) who experienced severe recurrent infections since 1 month of age. There is no family history of any similar disease. When the patient was 4 months old, she began receiving treatment with recombinant human G-CSF that resulted in a small increase in the neutrophil count sufficient for the prevention and treatment of bacterial infection. An analysis of complementary DNA for the patient's G-CSF receptor revealed a 3-base pair deletion in the juxtamembrane intracellular sequence. This deletion at the beginning of exon 16 was thought to be caused by alternative splicing; analysis of the DNA revealed a G-to-A point mutation of the final nucleotide of intron 15. To evaluate the functional activity of the G-CSF receptor with this 3-base pair deletion of the juxtamembrane region, we transfected this G-CSF receptor mutant into an interleukin 3-dependent cell line, BAF/3. BAF/3 cells expressing the mutant G-CSF receptor showed augmented proliferation activity in response to G-CSF compared with cells having the wild-type G-CSF receptor. Although the proliferation signal of G-CSF in normal hematopoiesis is transduced through the activation of MAP kinases, this G-CSF receptor mutant showed decreased activation of ERKI/2 in response to G-CSF compared with the wild type, but the transduced sig-nal for Stat3 activation by G-CSF was of the same magnitude as that of the wild-type G-CSF receptor. This result means that the augmented proliferation activity in response to G-CSF that we observed in cells having the G-CSF receptor gene with the 3-base pair deletion is transduced through an intracellular signaling pathway other than MAP kinase. Because SCN patients with a mutation in the G-CSF receptor frequently develop leukemia, this 3-base pair deletion in the juxtamembrane sequence of the G-CSF receptor gene in this patient may be one step in the course of leukemic transformation.
Yokoyama
Toshihiro
T
The First Department of Internal Medicine, Faculty of Medivine, Kyushu University, Fukuoka, Japan.
Okamura
Seiichi
S
Asano
Yoshinobu
Y
Kamezaki
Kenjirou
K
Numata
Akihiko
A
Kakumitsu
Haruko
H
Shide
Koutarou
K
Nakashima
Hitoshi
H
Taisuke
Kanaji
K
Sekine
Yuichi
Y
Mizuno
Yumi
Y
Okamura
Jun
J
Matsuda
Tadashi
T
Harada
Mine
M
Yoshiyuki
Niho
N
Shimoda
Kazuya
K
D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic; D002648:Child; D004247:DNA / Q000032:analysis; D005260:Female; D016368:Frameshift Mutation; D049690:History, Ancient; D006801:Humans; D007938:Leukemia / Q000235:genetics; D020935:MAP Kinase Signaling System; D009503:Neutropenia / Q000151:congenital* / Q000235:genetics*; D016188:Receptors, Granulocyte Colony-Stimulating Factor / Q000235:genetics*; D015398:Signal Transduction; D013577:Syndrome
D016188:Receptors, Granulocyte Colony-Stimulating Factor; D004247:DNA