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Amanda K. Arrington 1,*, Eileen L.

Heinrich 1, Wendy Lee 1, Marjun Duldulao 1,


Supriya Patel 2, Julian Sanchez 1, Julio Garcia-Aguilar 3 and Joseph Kim 1,*:
Prognostic and Predictive Roles of KRAS Mutation in
Colorectal Cancer, 2012, Int. J. Mol. Sci. 13, 12153-12168
Gen K-RAS

RAS adalah suatu kelompok gen yang berkaitan dengan protein yang mengkode membran-bound protein kelas 21
kD yang mengikat nukleotida guanin dan memiliki aktivitas GTPase intrinsik. Dua gen RAS yang pertama kali
ditemukan yaitu HRAS dan KRAS, gen ini diidentifikasi pada tahun 1975 dari penelitian terhadap dua virus
penyebab kanker, virus sarcoma Harvey dan virus sarcoma Kirsten, oleh Scolnick dkk. 1 Kemudian menyusul
ditemukan N-RAS yang ditemukan pada tahun 1982 dan ketiganya telah terbukti secara pasti terlibat dalam
patogenesis berbagai macam kanker. Dari tiga gen RAS yang diketahui, KRAS paling sering bermutasi pada kanker.

Gen KRAS mengkodekan protein asam amino 188 yang memiliki aktivitas katalitik. Translasi pasca modifikasi
protein K-RAS difasilitasi lokal ke dalam membran sel. Biasanya, protein K-RAS beraada dalam keadaan tidak aktif
didalam sel. K_RAS akan aktif saat reseptor transmembran terdekat (mis., growth factor receptors, G-protein
coupled receptors, toll-like receptors,, dll) Terikat oleh ligannya (Gambar 1). Siklus sinyal intraselular
berikutnya melibatkan guanine exchange factors (GEF) yang memfasilitasi pengaktifan RAS dengan mengganti
inactive GDP dengan GTP. Setelah diaktifkan, ras\akan memulai aktivasi downstream dari berbagai macam
efektor termasuk kinin serin / treonin, GTPase-activating proteins (GAPs), phosphoinositide 3-kinase
(PI3K), dan GEFs [11]. Ras akan dinonaktifkan saat molekul GTP diubah kembali menjadi molekul GDP [11]. Jika
KRAS bermutasi, K-RAS akan tetap berada pada fase GTP-bound protein. sehingga, pengaturan fungsi dari
downstream akan menghilang (
Seperti yang ditunjukkan oleh nama mereka, GEFs membantu pertukaran tersebut
dari PDB terikat dengan GTP, sedangkan GAP merangsang
kemampuan hidrolitik RAS untuk mengubah GTP terikat menjadi PDB.13
Lokalisasi membran yang tepat dan fungsinya
Protein RAS diatur oleh beberapa post-translational
Modifikasi pada motif "CAAX" C-terminal, termasuk
farnesilasi residu sistein, penghapusan proteolitik
dari terminal tiga residu (AAX), serta metilasi
dari residu sistein.15,19 Sebagai tambahan, membran plasma
Pelokalan KRAS juga membutuhkan poli-lisin dasar
daerah yang terletak tepat di hulu Cterminus.
19,20
Setelah benar lokal, KRAS menengahi segudang
peristiwa sinyal intraselular melalui berbagai efektor
jalur. Signaling oleh reseptor tirosin kinase (RTKs), di
khususnya reseptor faktor pertumbuhan epidermal (EGFR), adalah
model pembelajaran yang banyak digunakan dan dipahami dengan baik
Aktivasi KRAS (Gambar 1A) .16,21 Aktivasi EGFR atas
ligan mengikat dan fosforilasi auto berikutnya
membuat situs dok untuk protein adaptor growthfactor-
reseptor protein terikat 2 (GRB2), yang mengikat
Anak GEF dari Sevenless (SOS) di sitosol. Itu
perekrutan protein ini kompleks menjadi terfosforilasi
reseptor memungkinkan SOS berfungsi sebagai faktor pertukaran
KRAS, menghasilkan pertukaran nukleotida dan ikatan GTP
bentuk KRAS (Gambar 1A) .16,21,22

RAS proteins represent prototypical members of a large family of small GTP-binding proteins.13
The human RAS superfamily consists of more than 100 members, which can be
divided into six subfamilies.14 Three prototypical RAS proteins
include HRAS, NRAS, and KRAS.15 While they are highly
homologous in amino acid sequence and ubiquitously
expressed, KRAS is the only one that is essential for normal
development as shown by mouse genetic studies.16e18 KRAS
can be expressed as two different splice variants, referred to
as 4A and 4B, through alternative splicing within exon 4.15 The
4B variant is the dominant form commonly known as KRAS.8
KRAS is a membrane-bound GTPase that cycles between
an active GTP-bound form and an inactive GDP-bound form
due to the hydrolysis of the bound GTP (Fig. 1A).14,19 The
switches between these two states are controlled by two
classes of proteins: guanosine nucleotide exchange factors
(known as GEFs) and GTPase-activating proteins (known as
GAPs). As their names suggest, GEFs assist with the exchange
of bound GDP with GTP, whereas GAPs stimulate the
hydrolytic ability of RAS to convert bound GTP to GDP.13
The proper membrane localization and function of the
RAS proteins are regulated by several post-translational
modifications in the C-terminal CAAX motif, including
farnesylation of the cysteine residue, proteolytic removal
of the terminal three residues (AAX), as well as methylation
of the cysteine residue.15,19 In addition, the plasma membrane
localization of KRAS also requires a basic poly-lysine
region located immediately upstream of the Cterminus.
19,20
Once properly localized, KRAS mediates a myriad of
intracellular signaling events through its numerous effector
pathways. Signaling by receptor tyrosine kinases (RTKs), in
particular the epidermal growth factor receptor (EGFR), is
a widely-utilized and well-understood model for studying
KRAS activation (Fig. 1A).16,21 The activation of EGFR upon
ligand binding and its subsequent auto-phosphorylation
create a docking site for the adaptor protein growthfactor-
receptor-bound protein 2 (GRB2), which binds to
the GEF Son of Sevenless (SOS) in the cytosol. The
recruitment of this protein complex to the phosphorylated
receptor enables SOS to function as the exchange factor for
KRAS, resulting in nucleotide exchange and the GTP-bound
form of KRAS (Fig. 1A).16,21,22

RAS is the name given to a family of related genes that encode a class of 21 kD membrane-bound
proteins that bind guanine nucleotides and have intrinsic GTPase activity. The first two RAS
genes, HRAS and KRAS, were identified in 1975 from studies of two cancer-causing viruses, the
Harvey sarcoma virus and Kirsten sarcoma virus, by Scolnick et al. at the National Institutes of
Health (NIH) [10]. The human analog of this gene was subsequently discovered in 1982 and has
been intensely studied and implicated in the pathogenesis of many cancers. Of the three known
human RAS genes, KRAS is most frequently mutated in cancer [7].
The KRAS gene encodes a 188 amino acid protein that has inherent catalytic activity. Post
translation modification of this protein facilitates its localization to the cell membrane. Normally,
ras proteins exist in an inactive state in any given cell. All members of the ras family become
activated when a nearby transmembrane receptor (e.g., growth factor receptors, G-protein
coupled receptors, toll-like receptors, etc.) is bound by its corresponding ligand (Figure 1). The
subsequent intracellular signal cascade involves guanine exchange factors (GEF) which facilitate
the activation of ras by replacing the inactive GDP with GTP. Once activated, ras leads to the
downstream activation of a wide variety of effectors including serine/threonine kinases, GTPase-
activating proteins (GAPs), phosphoinositide 3-kinase (PI3K), and GEFs [11]. Ras is deactivated
when the GTP molecule is converted back to a GDP molecule [11]. If KRAS is mutated, it
remains in the GTP state. Therefore, KRAS remains in a constitutive GTP-bound state and, thus,
regulation of downstream functions is lost (Figure 2). For example, the dysregulated GTP-bound
activation of mutant-derived KRAS protein leads to unregulated downstream cell-growth.